http://journals.iucr.org/f/issues/2012/02/00/isscontsbdy.html Acta Crystallographica Section F: Structural Biology and Crystallization Communications is a rapid all-electronic journal, which provides a home for short communications on the crystallization and structure of biological macromolecules. Structures determined through structural genomics initiatives or from iterative studies such as those used in the pharmaceutical industry are particularly welcomed. Articles are available online when ready, making publication as fast as possible, and include unlimited free colour illustrations, movies and other enhancements. The editorial process is completely electronic with respect to deposition, submission, refereeing and publication. en Copyright (c) 2012 International Union of Crystallography 2012-02-01 International Union of Crystallography International Union of Crystallography http://journals.iucr.org urn:issn:1744-3091 Acta Crystallographica Section F: Structural Biology and Crystallization Communications is a rapid all-electronic journal, which provides a home for short communications on the crystallization and structure of biological macromolecules. Structures determined through structural genomics initiatives or from iterative studies such as those used in the pharmaceutical industry are particularly welcomed. Articles are available online when ready, making publication as fast as possible, and include unlimited free colour illustrations, movies and other enhancements. The editorial process is completely electronic with respect to deposition, submission, refereeing and publication. text/html Acta Crystallographica Section F: Structural Biology and Crystallization Communications, Volume 68, Part 2, 2012 text monthly 1 2002-01-01T00:00+00:00 2 68 2012-02-01 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications 115 urn:issn:1744-3091 med@iucr.org February 2012 2012-02-01 http://journals.iucr.org/logos/rss10f.gif http://journals.iucr.org/f/issues/2012/02/00/isscontsbdy.html Still image http://scripts.iucr.org/cgi-bin/paper?en5484 The crystal structures of tankyrase 1 (TNKS1) in complex with two small-molecule inhibitors, PJ34 and XAV939, both at 2.0 Å resolution, are reported. The structure of TNKS1 in complex with PJ34 reveals two molecules of PJ34 bound in the NAD+ donor pocket. One molecule is in the nicotinamide portion of the pocket, as previously observed in other PARP structures, while the second molecule is bound in the adenosine portion of the pocket. Additionally, unlike the unliganded crystallization system, the TNKS1–PJ34 crystallization system has the NAD+ donor site accessible to bulk solvent in the crystal, which allows displacement soaking. The TNKS1–PJ34 crystallization system was used to determine the structure of TNKS1 in complex with XAV939. These structures provide a basis for the start of a structure-based drug-design campaign for TNKS1. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Kirby, C.A. Cheung, A. Fazal, A. Shultz, M.D. Stams, T. 2012-01-21 doi:10.1107/S1744309111051219 International Union of Crystallography Crystal structures of human tankyrase I in complex with small-molecule inhibitors PJ34 and XAV939 have been determined, both to 2.0 Å. The TNKS1-PJ34 system allows for displacement soaking and, therefore, is an ideal system for structure-based drug design. EN tankyrase PARP domains inhibitor complexes displacement soaking The crystal structures of tankyrase 1 (TNKS1) in complex with two small-molecule inhibitors, PJ34 and XAV939, both at 2.0 Å resolution, are reported. The structure of TNKS1 in complex with PJ34 reveals two molecules of PJ34 bound in the NAD+ donor pocket. One molecule is in the nicotinamide portion of the pocket, as previously observed in other PARP structures, while the second molecule is bound in the adenosine portion of the pocket. Additionally, unlike the unliganded crystallization system, the TNKS1–PJ34 crystallization system has the NAD+ donor site accessible to bulk solvent in the crystal, which allows displacement soaking. The TNKS1–PJ34 crystallization system was used to determine the structure of TNKS1 in complex with XAV939. These structures provide a basis for the start of a structure-based drug-design campaign for TNKS1. text/html Structure of human tankyrase 1 in complex with small-molecule inhibitors PJ34 and XAV939 text 2 68 2012-01-21 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications structural communications 115 118 http://scripts.iucr.org/cgi-bin/paper?tb5045 The room-temperature (RT) X-ray structure of H/D-exchanged crambin is reported at 0.85 Å resolution. As one of the very few proteins refined with anisotropic atomic displacement parameters at two temperatures, the dynamics of atoms in the RT and 100 K structures are compared. Neutron diffraction data from an H/D-exchanged crambin crystal collected at the Protein Crystallography Station (PCS) showed diffraction beyond 1.1 Å resolution. This is the highest resolution neutron diffraction reported to date for a protein crystal and will reveal important details of the anisotropic motions of H and D atoms in protein structures. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Chen, J.C.-H. Fisher, Z. Kovalevsky, A.Y. Mustyakimov, M. Hanson, B.L. Zhurov, V.V. Langan, P. 2012-01-21 doi:10.1107/S1744309111051499 International Union of Crystallography The 0.85 Å room-temperature ultrahigh-resolution structure of H/D-exchanged crambin is reported. Preliminary 1.1 Å resolution neutron diffraction data have been collected at the neutron Protein Crystallography Station at LANSCE. EN crambin neutron diffraction ultrahigh resolution H/D exchange The room-temperature (RT) X-ray structure of H/D-exchanged crambin is reported at 0.85 Å resolution. As one of the very few proteins refined with anisotropic atomic displacement parameters at two temperatures, the dynamics of atoms in the RT and 100 K structures are compared. Neutron diffraction data from an H/D-exchanged crambin crystal collected at the Protein Crystallography Station (PCS) showed diffraction beyond 1.1 Å resolution. This is the highest resolution neutron diffraction reported to date for a protein crystal and will reveal important details of the anisotropic motions of H and D atoms in protein structures. text/html Room-temperature ultrahigh-resolution time-of-flight neutron and X-ray diffraction studies of H/D-exchanged crambin text 2 68 2012-01-21 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications structural communications 119 123 http://scripts.iucr.org/cgi-bin/paper?be5189 The X-ray structures of new crystal forms of peptidyl-tRNA hydrolase from M. tuberculosis reported here and the results of previous X-ray studies of the enzyme from different sources provide a picture of the functionally relevant plasticity of the protein molecule. The new X-ray results confirm the connection deduced previously between the closure of the lid at the peptide-binding site and the opening of the gate that separates the peptide-binding and tRNA-binding sites. The plasticity of the molecule indicated by X-ray structures is in general agreement with that deduced from the available solution NMR results. The correlation between the lid and the gate movements is not, however, observed in the NMR structure. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Selvaraj, M. Ahmad, R. Varshney, U. Vijayan, M. 2012-01-21 doi:10.1107/S1744309111052341 International Union of Crystallography The structures of new crystal forms of M. tuberculosis peptidyl-tRNA hydrolase confirm and provide further elaboration of the functionally relevant plasticity of the enzyme molecule. EN enzyme action peptidyl-tRNA binding site protein synthesis molecular plasticity crystal and solution structures The X-ray structures of new crystal forms of peptidyl-tRNA hydrolase from M. tuberculosis reported here and the results of previous X-ray studies of the enzyme from different sources provide a picture of the functionally relevant plasticity of the protein molecule. The new X-ray results confirm the connection deduced previously between the closure of the lid at the peptide-binding site and the opening of the gate that separates the peptide-binding and tRNA-binding sites. The plasticity of the molecule indicated by X-ray structures is in general agreement with that deduced from the available solution NMR results. The correlation between the lid and the gate movements is not, however, observed in the NMR structure. text/html Structures of new crystal forms of Mycobacterium tuberculosis peptidyl-tRNA hydrolase and functionally important plasticity of the molecule text 2 68 2012-01-21 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications structural communications 124 128 http://scripts.iucr.org/cgi-bin/paper?hv5208 Pectin methylesterases (PMEs) are family 8 carbohydrate esterases (CE8s) which remove the methyl group from methylesterified galacturonic acid (GalA) residues within pectin. Although the role of pectinases such as PMEs within dedicated phytopathogens has been well established, the significance of homologous enzymes found within the genomes of human enteropathogens remains to be determined. Presented here is the low-resolution (3.5 Å) structure of the CE8 from Yersinia enterocolitica (YeCE8). The high degree of structural conservation in the topology of the active-site cleft and catalytic apparatus that is shared with a characterized PME from a bacterial phytopathogen (i) indicates that YeCE8 is active on methylated pectin and (ii) highlights a more prominent role for pectin utilization in Yersinia than in other enteropathogenic species. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Boraston, A.B. Abbott, D.W. 2012-01-21 doi:10.1107/S1744309111055400 International Union of Crystallography The first crystal structure of a pectin methylesterase from an enteropathogen is presented. This enzyme from Y. enterocolitica has biological significance for the evolution of pectin-metabolic pathways within pectinolytic bacteria and related agents of foodborne illness. EN carbohydrate esterases pectins methylesterification enteropathogens Pectin methylesterases (PMEs) are family 8 carbohydrate esterases (CE8s) which remove the methyl group from methylesterified galacturonic acid (GalA) residues within pectin. Although the role of pectinases such as PMEs within dedicated phytopathogens has been well established, the significance of homologous enzymes found within the genomes of human enteropathogens remains to be determined. Presented here is the low-resolution (3.5 Å) structure of the CE8 from Yersinia enterocolitica (YeCE8). The high degree of structural conservation in the topology of the active-site cleft and catalytic apparatus that is shared with a characterized PME from a bacterial phytopathogen (i) indicates that YeCE8 is active on methylated pectin and (ii) highlights a more prominent role for pectin utilization in Yersinia than in other enteropathogenic species. text/html Structure of a pectin methylesterase from Yersinia enterocolitica text 2 68 2012-01-21 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications structural communications 129 133 http://scripts.iucr.org/cgi-bin/paper?tb5044 A new crystal lattice structure of Helicobacter pylori neutrophil-activating protein (HP-NAP) has been determined in two forms: the native state (Apo) at 2.20 Å resolution and an iron-loaded form (Fe-load) at 2.50 Å resolution. The highly solvated packing of the dodecameric shell is suitable for crystallographic study of the metal ion-uptake pathway. Like other bacterioferritins, HP-NAP forms a spherical dodecamer with 23 symmetry including two kinds of channels. Iron loading causes a series of conformational changes of amino-acid residues (Trp26, Asp52 and Glu56) at the ferroxidase centre. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Tsuruta, O. Yokoyama, H. Fujii, S. 2012-01-25 doi:10.1107/S1744309111052675 International Union of Crystallography A new crystal lattice structure of H. pylori neutrophil-activating protein has been determined. Iron loading causes a series of conformational changes at the ferroxidase centre. EN Helicobacter pylori neutrophil-activating proteins ferroxidase centre solvent channels pores A new crystal lattice structure of Helicobacter pylori neutrophil-activating protein (HP-NAP) has been determined in two forms: the native state (Apo) at 2.20 Å resolution and an iron-loaded form (Fe-load) at 2.50 Å resolution. The highly solvated packing of the dodecameric shell is suitable for crystallographic study of the metal ion-uptake pathway. Like other bacterioferritins, HP-NAP forms a spherical dodecamer with 23 symmetry including two kinds of channels. Iron loading causes a series of conformational changes of amino-acid residues (Trp26, Asp52 and Glu56) at the ferroxidase centre. text/html A new crystal lattice structure of Helicobacter pylori neutrophil-activating protein (HP-NAP) text 2 68 2012-01-25 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications structural communications 134 140 http://scripts.iucr.org/cgi-bin/paper?tb5041 Thermostable enzymes employ various structural features dictated at the amino-acid sequence level that allow them to maintain their integrity at higher temperatures. Many hypotheses as to the nature of thermal stability have been proposed, including optimized core hydrophobicity and an increase in charged surface residues to enhance polar solvent interactions for solubility. Here, the three-dimensional structure of the family GH11 xylanase from the moderate thermophile Thermobifida fusca in its trapped covalent glycosyl-enzyme intermediate complex is presented. Interactions with the bound ligand show fewer direct hydrogen bonds from ligand to protein than observed in previous complexes from other species and imply that binding of the xylan substrate involves several water-mediated hydrogen bonds. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Lammerts van Bueren, A. Otani, S. Friis, E.P. Wilson, K.S. Davies, G.J. 2012-01-25 doi:10.1107/S1744309111049608 International Union of Crystallography The structure of a thermostable xylanase from T. fusca is reported. EN xylanases glycoside hydrolases covalent intermediate thermostability Thermostable enzymes employ various structural features dictated at the amino-acid sequence level that allow them to maintain their integrity at higher temperatures. Many hypotheses as to the nature of thermal stability have been proposed, including optimized core hydrophobicity and an increase in charged surface residues to enhance polar solvent interactions for solubility. Here, the three-dimensional structure of the family GH11 xylanase from the moderate thermophile Thermobifida fusca in its trapped covalent glycosyl-enzyme intermediate complex is presented. Interactions with the bound ligand show fewer direct hydrogen bonds from ligand to protein than observed in previous complexes from other species and imply that binding of the xylan substrate involves several water-mediated hydrogen bonds. text/html Three-dimensional structure of a thermophilic family GH11 xylanase from Thermobifida fusca text 2 68 2012-01-25 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications structural communications 141 144 http://scripts.iucr.org/cgi-bin/paper?wd5165 The crystal structure of the interleukin-2 tyrosine kinase Src homology domain (Itk SH2) is described and it is found that unlike in studies of this domain using NMR spectroscopy, cis–trans-prolyl isomerization is not readily detected in the crystal structure. Based on similarities between the Itk SH2 crystal form and the cis form of the Itk SH2 NMR structure, it is concluded that it is likely that the prolyl imide bond at least in part adopts the cis conformation in the crystal form. However, the lack of high-resolution data and the dynamic nature of the proline-containing loop mean that the precise imide-bond conformation cannot be determined and prolyl cis–trans isomerization in the crystal cannot be ruled out. Given the preponderance of structures that have been solved by X-ray crystallography in the Protein Data Bank, this result supports the notion that prolyl isomerization in folded proteins has been underestimated among known structures. Interestingly, while the precise status of the proline residue is ambiguous, Itk SH2 crystallizes as a domain-swapped dimer. The domain-swapped structure of Itk SH2 is similar to the domain-swapped SH2 domains of Grb2 and Nck, with domain swapping occurring at the β-meander region of all three SH2 domains. Thus, for Itk SH2 structural analysis by NMR spectroscopy and X-ray crystallography revealed very different structural features: proline isomerization versus domain-swapped dimerization, respectively. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Joseph, R.E. Ginder, N.D. Hoy, J.A. Nix, J.C. Fulton, D.B. Honzatko, R.B. Andreotti, A.H. 2012-01-25 doi:10.1107/S1744309111049761 International Union of Crystallography The interleukin-2 tyrosine kinase Src homology 2 domain was crystallized and its structure was solved to 2.35 Å resolution. The structure reveals a domain-swapped dimer that is related to other dimeric SH2 domains solved previously. The cis–trans-prolyl isomerization that is evident from solution studies of Itk SH2 cannot be observed in the crystal structure. EN IL-2-inducible T-cell kinase Src homology 2 domain proline isomerization domain swapping The crystal structure of the interleukin-2 tyrosine kinase Src homology domain (Itk SH2) is described and it is found that unlike in studies of this domain using NMR spectroscopy, cis–trans-prolyl isomerization is not readily detected in the crystal structure. Based on similarities between the Itk SH2 crystal form and the cis form of the Itk SH2 NMR structure, it is concluded that it is likely that the prolyl imide bond at least in part adopts the cis conformation in the crystal form. However, the lack of high-resolution data and the dynamic nature of the proline-containing loop mean that the precise imide-bond conformation cannot be determined and prolyl cis–trans isomerization in the crystal cannot be ruled out. Given the preponderance of structures that have been solved by X-ray crystallography in the Protein Data Bank, this result supports the notion that prolyl isomerization in folded proteins has been underestimated among known structures. Interestingly, while the precise status of the proline residue is ambiguous, Itk SH2 crystallizes as a domain-swapped dimer. The domain-swapped structure of Itk SH2 is similar to the domain-swapped SH2 domains of Grb2 and Nck, with domain swapping occurring at the β-meander region of all three SH2 domains. Thus, for Itk SH2 structural analysis by NMR spectroscopy and X-ray crystallography revealed very different structural features: proline isomerization versus domain-swapped dimerization, respectively. text/html Structure of the interleukin-2 tyrosine kinase Src homology 2 domain; comparison between X-ray and NMR-derived structures text 2 68 2012-01-25 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications structural communications 145 153 http://scripts.iucr.org/cgi-bin/paper?bw5401 Crystallization of contaminating proteins is a frequently encountered problem for macromolecular crystallographers. In this study, an attempt was made to obtain a binary cocrystal structure of the SH3 domain of cortactin and a 17-residue peptide from the Arg nonreceptor tyrosine kinase encompassing a PxxPxxPxxP (PxxP1) motif. However, cocrystals could only be obtained in the presence of trace amounts of a contaminating protein. A structure solution obtained by molecular replacement followed by ARP/wARP automatic model building allowed a `sequence-by-crystallography' approach to discover that the contaminating protein was lysozyme. This 1.65 Å resolution crystal structure determination of a 1:1:1 heterotrimeric complex of Arg, cortactin and lysozyme thus provides an unusual `caveat emptor' warning of the dangers that underpurified proteins harbor for macromolecular crystallographers. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Liu, W. MacGrath, S.M. Koleske, A.J. Boggon, T.J. 2012-01-25 doi:10.1107/S1744309111056132 International Union of Crystallography An unusual case of trace amounts of a contaminating protein facilitating formation of a heterotrimeric protein complex. EN cortactin Arg protein–protein complexes cytoskeleton Crystallization of contaminating proteins is a frequently encountered problem for macromolecular crystallographers. In this study, an attempt was made to obtain a binary cocrystal structure of the SH3 domain of cortactin and a 17-residue peptide from the Arg nonreceptor tyrosine kinase encompassing a PxxPxxPxxP (PxxP1) motif. However, cocrystals could only be obtained in the presence of trace amounts of a contaminating protein. A structure solution obtained by molecular replacement followed by ARP/wARP automatic model building allowed a `sequence-by-crystallography' approach to discover that the contaminating protein was lysozyme. This 1.65 Å resolution crystal structure determination of a 1:1:1 heterotrimeric complex of Arg, cortactin and lysozyme thus provides an unusual `caveat emptor' warning of the dangers that underpurified proteins harbor for macromolecular crystallographers. text/html Lysozyme contamination facilitates crystallization of a heterotrimeric cortactin–Arg–lysozyme complex text 2 68 2012-01-25 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications structural communications 154 158 http://scripts.iucr.org/cgi-bin/paper?pu5348 PACSIN-family proteins are cytoplasmic proteins that have vesicle-transport, membrane-dynamics, actin-reorganization and microtubule activities. Here, the N-terminal F-BAR domain of mouse PACSIN 3, which contains 341 amino acids, was successfully cloned, purified and crystallized. The crystal of PACSIN 3 (1–341) diffracted to 2.6 Å resolution and belonged to space group P21, with unit-cell parameters a = 46.9, b = 54.7, c = 193.7 Å, α = 90, β = 96.9, γ = 90°. These data should provide further information on PACSIN-family protein structures. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Bai, X. Meng, G. Zheng, X. 2012-01-25 doi:10.1107/S1744309111049116 International Union of Crystallography The N-terminal F-BAR domain of mouse PACSIN 3, which contains 341 amino acids, has been successfully cloned, purified and crystallized. EN PACSIN 3 BAR-domain proteins F-BAR domain PACSIN-family proteins are cytoplasmic proteins that have vesicle-transport, membrane-dynamics, actin-reorganization and microtubule activities. Here, the N-terminal F-BAR domain of mouse PACSIN 3, which contains 341 amino acids, was successfully cloned, purified and crystallized. The crystal of PACSIN 3 (1–341) diffracted to 2.6 Å resolution and belonged to space group P21, with unit-cell parameters a = 46.9, b = 54.7, c = 193.7 Å, α = 90, β = 96.9, γ = 90°. These data should provide further information on PACSIN-family protein structures. text/html Cloning, purification, crystallization and preliminary X-ray diffraction analysis of mouse PACSIN 3 protein text 2 68 2012-01-25 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 159 162 http://scripts.iucr.org/cgi-bin/paper?ub5027 DNA ligases join single-strand breaks in double-stranded DNA by catalyzing the formation of a phosphodiester bond between adjacent 5′-phosphate and 3′-hydroxyl termini. Their function is essential to maintain the integrity of the genome in DNA replication, recombination and repair. A recombinant ATP-dependent DNA ligase from the hyperthermophilic anaerobic archaeon Thermococcus sibiricus was expressed in Escherichia coli and purified. Crystals were grown by vapour diffusion using the hanging-drop method with 17%(w/v) PEG 4000 and 8.5%(v/v) 2-propanol as precipitants. A diffraction experiment was performed with a single crystal, which diffracted X-rays to 3.0 Å resolution. The crystal belonged to space group P212121, with unit-cell parameters a = 58.590, b = 87.540, c = 126.300 Å. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Petrova, T.E. Bezsudnova, E.Y. Dorokhov, B.D. Slutskaya, E.S. Polyakov, K.M. Dorovatovskiy, P.V. Ravin, N.V. Skryabin, K.G. Kovalchuk, M.V. Popov, V.O. 2012-01-25 doi:10.1107/S1744309111050913 International Union of Crystallography A recombinant ATP-dependent DNA ligase from the hyperthermophilic anaerobic archaeon Thermococcus sibiricus was expressed in Escherichia coli and purified. Crystals were grown by vapour diffusion using the hanging-drop method. EN DNA ligases archaea DNA ligases join single-strand breaks in double-stranded DNA by catalyzing the formation of a phosphodiester bond between adjacent 5′-phosphate and 3′-hydroxyl termini. Their function is essential to maintain the integrity of the genome in DNA replication, recombination and repair. A recombinant ATP-dependent DNA ligase from the hyperthermophilic anaerobic archaeon Thermococcus sibiricus was expressed in Escherichia coli and purified. Crystals were grown by vapour diffusion using the hanging-drop method with 17%(w/v) PEG 4000 and 8.5%(v/v) 2-propanol as precipitants. A diffraction experiment was performed with a single crystal, which diffracted X-rays to 3.0 Å resolution. The crystal belonged to space group P212121, with unit-cell parameters a = 58.590, b = 87.540, c = 126.300 Å. text/html Expression, purification, crystallization and preliminary crystallographic analysis of a thermostable DNA ligase from the archaeon Thermococcus sibiricus text 2 68 2012-01-25 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 163 165 http://scripts.iucr.org/cgi-bin/paper?xb5043 Bacteriophage T7 attaches to its host using the C-terminal domains of its six fibres, which are trimers of the gp17 protein. A C-terminal fragment of gp17 consisting of amino acids 371–553 has been expressed, purified and crystallized. Crystals of two forms were obtained, belonging to space group P212121 (unit-cell parameters a = 61.2, b = 86.0, c = 118.4 Å) and space group C2221 (unit-cell parameters a = 68.3, b = 145.6, c = 172.1 Å). They diffracted to 1.9 and 2.0 Å resolution, respectively. Both crystals are expected to contain one trimer in the asymmetric unit. Multiwavelength anomalous dispersion phasing with a mercury derivative is in progress. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Garcia-Doval, C. van Raaij, M.J. 2012-01-25 doi:10.1107/S1744309111051049 International Union of Crystallography The C-terminal domain of the bacteriophage T7 fibre protein gp17, consisting of amino acids 371–553, has been crystallized. Diffraction data have been obtained to around 2.0 Å resolution from two different crystal forms. Multiwavelength anomalous dispersion phasing with a mercury derivative is in progress. EN bacteriophage T7 gp17 protein Bacteriophage T7 attaches to its host using the C-terminal domains of its six fibres, which are trimers of the gp17 protein. A C-terminal fragment of gp17 consisting of amino acids 371–553 has been expressed, purified and crystallized. Crystals of two forms were obtained, belonging to space group P212121 (unit-cell parameters a = 61.2, b = 86.0, c = 118.4 Å) and space group C2221 (unit-cell parameters a = 68.3, b = 145.6, c = 172.1 Å). They diffracted to 1.9 and 2.0 Å resolution, respectively. Both crystals are expected to contain one trimer in the asymmetric unit. Multiwavelength anomalous dispersion phasing with a mercury derivative is in progress. text/html Crystallization of the C-terminal domain of the bacteriophage T7 fibre protein gp17 text 2 68 2012-01-25 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 166 171 http://scripts.iucr.org/cgi-bin/paper?rl5014 The diphtheria toxin repressor (DtxR) is a metal-ion-dependent transcriptional regulator which regulates genes encoding proteins involved in metal-ion uptake to maintain metal-ion homeostasis. DtxR from Thermoplasma acidophilum was cloned and overexpressed in Escherichia coli. Crystals of N-terminally His-tagged DtxR were obtained by hanging-drop vapour diffusion and diffracted to 1.8 Å resolution. DtxR was crystallized at 296 K using polyethylene glycol 4000 as a precipitant. The crystals belonged to the orthorhombic space group P21212, with unit-cell parameters a = 61.14, b = 84.61, c = 46.91 Å, α = β = γ = 90°. The asymmetric unit contained approximately one monomer of DtxR, giving a crystal volume per mass (VM) of 2.22 Å3 Da−1 and a solvent content of 44.6%. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Yeo, H.K. Kang, J. Park, Y.W. Sung, J.-S. Lee, J.Y. 2012-01-26 doi:10.1107/S1744309111051700 International Union of Crystallography Orthorhombic crystals of DtxR from T. acidophilum have been obtained. X-ray data were collected to 1.8 Å resolution using synchrotron radiation. EN DtxR transcriptional regulators metalloregulatory proteins Thermoplasma acidophilum The diphtheria toxin repressor (DtxR) is a metal-ion-dependent transcriptional regulator which regulates genes encoding proteins involved in metal-ion uptake to maintain metal-ion homeostasis. DtxR from Thermoplasma acidophilum was cloned and overexpressed in Escherichia coli. Crystals of N-terminally His-tagged DtxR were obtained by hanging-drop vapour diffusion and diffracted to 1.8 Å resolution. DtxR was crystallized at 296 K using polyethylene glycol 4000 as a precipitant. The crystals belonged to the orthorhombic space group P21212, with unit-cell parameters a = 61.14, b = 84.61, c = 46.91 Å, α = β = γ = 90°. The asymmetric unit contained approximately one monomer of DtxR, giving a crystal volume per mass (VM) of 2.22 Å3 Da−1 and a solvent content of 44.6%. text/html Crystallization and preliminary X-ray diffraction analysis of the metalloregulatory protein DtxR from Thermoplasma acidophilum text 2 68 2012-01-26 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 172 174 http://scripts.iucr.org/cgi-bin/paper?bo5096 LipC12, a true lipase from family I.1 of bacterial lipases which was previously isolated through a metagenomics approach, contains 293 amino acids. Among lipases of known three-dimensional structure, it has a sequence identity of 47% to the lipase from Pseudomonas aeruginosa PAO1. Recombinant N-terminally His6-tagged LipC12 protein was expressed in Escherichia coli, purified in a homogenous form and crystallized in several conditions, with the best crystals being obtained using 2.0 M sodium formate and 0.1 M bis-tris propane pH 7.0. X-ray diffraction data were collected to 2.70 Å resolution. The crystals belonged to the tetragonal space group P4122, with unit-cell parameters a = b = 58.62, c = 192.60 Å. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Martini, V.P. Glogauer, A. Iulek, J. Souza, E.M. Pedrosa, F.O. Krieger, N. 2012-01-26 doi:10.1107/S1744309111051323 International Union of Crystallography The preliminary X-ray analysis of LipC12, the first lipase isolated through a metagenomics approach to be crystallized, is reported. EN LipC12 lipases metagenomics LipC12, a true lipase from family I.1 of bacterial lipases which was previously isolated through a metagenomics approach, contains 293 amino acids. Among lipases of known three-dimensional structure, it has a sequence identity of 47% to the lipase from Pseudomonas aeruginosa PAO1. Recombinant N-terminally His6-tagged LipC12 protein was expressed in Escherichia coli, purified in a homogenous form and crystallized in several conditions, with the best crystals being obtained using 2.0 M sodium formate and 0.1 M bis-tris propane pH 7.0. X-ray diffraction data were collected to 2.70 Å resolution. The crystals belonged to the tetragonal space group P4122, with unit-cell parameters a = b = 58.62, c = 192.60 Å. text/html Crystallization and preliminary crystallographic analysis of LipC12, a true lipase isolated through a metagenomics approach text 2 68 2012-01-26 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 175 177 http://scripts.iucr.org/cgi-bin/paper?fw5343 Mycobacterium tuberculosis DNA gyrase, a nanomachine involved in regulation of DNA topology, is the only type II topoisomerase present in this organism and hence is the sole target of fluoroquinolone in the treatment of tuberculosis. The C-terminal domain (CTD) of the DNA gyrase A subunit possesses a unique feature, the ability to wrap DNA in a chiral manner, that plays an essential role during the catalytic cycle. A construct of 36 kDa corresponding to this domain has been overproduced, purified and crystallized. Diffraction data were collected to 1.55 Å resolution. Cleavage of the N-terminal His tag was crucial for obtaining crystals. The crystals belonged to space group P212121, with one molecule in the asymmetric unit and a low solvent content (33%). This is the first report of the crystallization and preliminary X-ray diffraction studies of a DNA gyrase CTD from a species that contains one unique type II topoisomerase. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Darmon, A. Piton, J. Roué, M. Petrella, S. Aubry, A. Mayer, C. 2012-01-26 doi:10.1107/S1744309111051888 International Union of Crystallography The M. tuberculosis DNA gyrase A C-terminal domain (CTD) was crystallized using the hanging-drop vapour-diffusion method. The crystals belonged to space group P212121 and diffraction data were collected to a resolution of 1.55 Å. EN tuberculosis Mycobacterium tuberculosis type II topoisomerases DNA gyrases C-terminal domain Mycobacterium tuberculosis DNA gyrase, a nanomachine involved in regulation of DNA topology, is the only type II topoisomerase present in this organism and hence is the sole target of fluoroquinolone in the treatment of tuberculosis. The C-terminal domain (CTD) of the DNA gyrase A subunit possesses a unique feature, the ability to wrap DNA in a chiral manner, that plays an essential role during the catalytic cycle. A construct of 36 kDa corresponding to this domain has been overproduced, purified and crystallized. Diffraction data were collected to 1.55 Å resolution. Cleavage of the N-terminal His tag was crucial for obtaining crystals. The crystals belonged to space group P212121, with one molecule in the asymmetric unit and a low solvent content (33%). This is the first report of the crystallization and preliminary X-ray diffraction studies of a DNA gyrase CTD from a species that contains one unique type II topoisomerase. text/html Purification, crystallization and preliminary X-ray crystallographic studies of the Mycobacterium tuberculosis DNA gyrase CTD text 2 68 2012-01-26 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 178 180 http://scripts.iucr.org/cgi-bin/paper?pu5351 The production of the PEL polysaccharide in Pseudomonas aeruginosa requires the binding of bis-(3′,5′)-cyclic dimeric guanosine monophosphate (c-di-GMP) to the cytoplasmic GGDEF domain of the inner membrane protein PelD. Here, the overexpression, purification and crystallization of a soluble construct of PelD that encompasses the GGDEF domain and a predicted GAF domain is reported. Diffraction-quality crystals were grown using the hanging-drop vapour-diffusion method. The crystals grew as flat plates, with unit-cell parameters a = 88.3, b = 114.0, c = 61.9 Å, α = β = γ = 90.0°. The PelD crystals exhibited the symmetry of space group P21212 and diffracted to a minimum d-spacing of 2.2 Å. On the basis of the Matthews coefficient (VM = 2.29 Å3 Da−1), it was estimated that two molecules are present in the asymmetric unit. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Marmont, L.S. Whitney, J.C. Robinson, H. Colvin, K.M. Parsek, M.R. Howell, P.L. 2012-01-26 doi:10.1107/S1744309111052109 International Union of Crystallography The expression, purification and crystallization of a soluble construct of the putative inner membrane protein PelD from P. aeruginosa is described. The crystals diffracted to a resolution of 2.2 Å. EN PelD pellicles Pseudomonas aeruginosa inner membrane proteins cystic fibrosis biofilms exopolysaccharides c-di-GMP receptors GGDEF domains GAF domains The production of the PEL polysaccharide in Pseudomonas aeruginosa requires the binding of bis-(3′,5′)-cyclic dimeric guanosine monophosphate (c-di-GMP) to the cytoplasmic GGDEF domain of the inner membrane protein PelD. Here, the overexpression, purification and crystallization of a soluble construct of PelD that encompasses the GGDEF domain and a predicted GAF domain is reported. Diffraction-quality crystals were grown using the hanging-drop vapour-diffusion method. The crystals grew as flat plates, with unit-cell parameters a = 88.3, b = 114.0, c = 61.9 Å, α = β = γ = 90.0°. The PelD crystals exhibited the symmetry of space group P21212 and diffracted to a minimum d-spacing of 2.2 Å. On the basis of the Matthews coefficient (VM = 2.29 Å3 Da−1), it was estimated that two molecules are present in the asymmetric unit. text/html Expression, purification, crystallization and preliminary X-ray analysis of Pseudomonas aeruginosa PelD text 2 68 2012-01-26 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 181 184 http://scripts.iucr.org/cgi-bin/paper?pu5353 Eukaryotic poly(A)-binding protein (PABP) commonly binds to the 3′-UTR poly(A) tail of every mRNA, but it also binds to the 5′-UTR of PABP mRNA for autoregulation of its expression. In the sequence of the latter binding site, the contiguous A residues are segmented discretely by the insertion of short pyrimidine oligonucleotides as linkers, so that (A)6–8 segments are repeated six times. This differs from the poly(A)-tail sequence, which has a higher binding affinity for PABP. In order to examine whether the A-rich repeats have a functional structure, several RNA/DNA analogues were subjected to crystallization. It was found that some of them could be crystallized. Single crystals thus obtained diffracted to 4.1 Å resolution. The fact that the repeated sequences can be crystallized suggests the possibility that the autoregulatory sequence in PABP mRNA has a specific structure which impedes the binding of PABP. When PABP is excessively produced, it could bind to this sequence by releasing the structure in order to interfere with initiation-complex formation for suppression of PABP translation. Otherwise, PABP at low concentration preferentially binds to the poly(A) tail of PABP mRNA. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Kikuchi, K. Shimizu, S. Sato, Y. Morishita, E.C. Takénaka, A. 2012-01-26 doi:10.1107/S1744309111052110 International Union of Crystallography Several DNA and RNA fragments containing tandem AAAAAA repeats, which were synthesized as analogues of the A-rich repeats found in the autoregulatory sequence of PABP mRNA, have been crystallized, suggesting the possibility that the autoregulatory sequence has a specific structure which impedes the binding of PABP. EN A-rich repeats PABP mRNA PABP autoregulatory sequences Eukaryotic poly(A)-binding protein (PABP) commonly binds to the 3′-UTR poly(A) tail of every mRNA, but it also binds to the 5′-UTR of PABP mRNA for autoregulation of its expression. In the sequence of the latter binding site, the contiguous A residues are segmented discretely by the insertion of short pyrimidine oligonucleotides as linkers, so that (A)6–8 segments are repeated six times. This differs from the poly(A)-tail sequence, which has a higher binding affinity for PABP. In order to examine whether the A-rich repeats have a functional structure, several RNA/DNA analogues were subjected to crystallization. It was found that some of them could be crystallized. Single crystals thus obtained diffracted to 4.1 Å resolution. The fact that the repeated sequences can be crystallized suggests the possibility that the autoregulatory sequence in PABP mRNA has a specific structure which impedes the binding of PABP. When PABP is excessively produced, it could bind to this sequence by releasing the structure in order to interfere with initiation-complex formation for suppression of PABP translation. Otherwise, PABP at low concentration preferentially binds to the poly(A) tail of PABP mRNA. text/html Crystallization of oligonucleotides containing A-rich repeats suggests a structural contribution to the autoregulation mechanism of PABP translation text 2 68 2012-01-26 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 185 189 http://scripts.iucr.org/cgi-bin/paper?ub5028 Baculovirus envelope protein ODV-E66 (67–704), in which the N-terminal 66 amino acids are truncated, is a chondroitin lyase. It digests chondroitin and chondroitin 6-sulfate efficiently, but does not digest chondroitin 4-sulfate. This unique characteristic is useful for the preparation of specific chondroitin oligosaccharides and for investigation of the mechanism of baculovirus infection. ODV-E66 (67–704) was crystallized; the crystal diffracted to 1.8 Å resolution and belonged to space group P62 or P64, with unit-cell parameters a = b = 113.5, c = 101.5 Å. One molecule is assumed to be present per asymmetric unit, which gives a Matthews coefficient of 2.54 Å3 Da−1. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Kawaguchi, Y. Sugiura, N. Onishi, M. Kimata, K. Kimura, M. Kakuta, Y. 2012-01-26 doi:10.1107/S1744309111053164 International Union of Crystallography Baculovirus envelope protein ODV-E66 (67–704) was crystallized; the crystal diffracted to 1.8 Å resolution and belonged to space group P62 or P64, with unit-cell parameters a = b = 113.5, c = 101.5 Å. EN chondroitinases chondroitin sulfate lyases polysaccharide lyase family 8 Baculovirus envelope protein ODV-E66 (67–704), in which the N-terminal 66 amino acids are truncated, is a chondroitin lyase. It digests chondroitin and chondroitin 6-sulfate efficiently, but does not digest chondroitin 4-sulfate. This unique characteristic is useful for the preparation of specific chondroitin oligosaccharides and for investigation of the mechanism of baculovirus infection. ODV-E66 (67–704) was crystallized; the crystal diffracted to 1.8 Å resolution and belonged to space group P62 or P64, with unit-cell parameters a = b = 113.5, c = 101.5 Å. One molecule is assumed to be present per asymmetric unit, which gives a Matthews coefficient of 2.54 Å3 Da−1. text/html Crystallization and X-ray diffraction analysis of chondroitin lyase from baculovirus: envelope protein ODV-E66 text 2 68 2012-01-26 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 190 192 http://scripts.iucr.org/cgi-bin/paper?bo5098 Dioscorin, the major tuber storage protein in yam, has been reported to possess carbonic anhydrase, trypsin inhibitor, angiotensin-converting enzyme (ACE) inhibitor, free-radical scavenger, dehydroascorbate reductase and monodehydroascorbate reductase activities. Recent research has also found that dioscorin can enhance immune modulation via the toll-like receptor 4 (TLR-4) signal transduction pathway in RAW 264.7 cells, murine bone-marrow cells and human monocytes ex vivo. Resolving the structure of dioscorin would help in better understanding its activities and would provide clues to understanding the mechanism of its multiple functions. The full-length protein (residues 1–246) with an additional His6 tag at the N-terminus was expressed in Escherichia coli Rosetta (DE3) cells. After His-tag cleavage and purification, the protein was crystallized by the sitting-drop vapour-diffusion method at 278 K. An X-ray diffraction data set was collected to a resolution of 2.11 Å using a synchrotron X-ray source. The crystal belonged to space group C2221, with unit-cell parameters a = 83.5, b = 156.8, c = 83.6 Å, and was estimated to contain two protein molecules per asymmetric unit. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Xue, Y.-L. Miyakawa, T. Sawano, Y. Tanokura, M. 2012-01-26 doi:10.1107/S1744309111053723 International Union of Crystallography Dioscorin from D. japonica was expressed, purified and crystallized using the sitting-drop vapour-diffusion method. The dioscorin crystal diffracted X-rays to 2.11 Å resolution. EN dioscorin Dioscorea japonica Dioscorin, the major tuber storage protein in yam, has been reported to possess carbonic anhydrase, trypsin inhibitor, angiotensin-converting enzyme (ACE) inhibitor, free-radical scavenger, dehydroascorbate reductase and monodehydroascorbate reductase activities. Recent research has also found that dioscorin can enhance immune modulation via the toll-like receptor 4 (TLR-4) signal transduction pathway in RAW 264.7 cells, murine bone-marrow cells and human monocytes ex vivo. Resolving the structure of dioscorin would help in better understanding its activities and would provide clues to understanding the mechanism of its multiple functions. The full-length protein (residues 1–246) with an additional His6 tag at the N-terminus was expressed in Escherichia coli Rosetta (DE3) cells. After His-tag cleavage and purification, the protein was crystallized by the sitting-drop vapour-diffusion method at 278 K. An X-ray diffraction data set was collected to a resolution of 2.11 Å using a synchrotron X-ray source. The crystal belonged to space group C2221, with unit-cell parameters a = 83.5, b = 156.8, c = 83.6 Å, and was estimated to contain two protein molecules per asymmetric unit. text/html Crystallization and preliminary X-ray crystallographic analysis of dioscorin from Dioscorea japonica text 2 68 2012-01-26 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 193 195 http://scripts.iucr.org/cgi-bin/paper?rl5015 Arylamine N-acetyltransferases (NATs) are xenobiotic metabolizing enzymes (XMEs) that catalyze the acetylation of arylamines. All functional NATs described to date possess a strictly conserved Cys-His-Asp catalytic triad. Here, the purification, crystallization and preliminary X-ray characterization of Bacillus cereus arylamine N-acetyltransferase 3 [(BACCR)NAT3], a putative NAT isoenzyme that possesses a unique catalytic triad containing a glutamate residue, is reported. The crystal diffracted to 2.42 Å resolution and belonged to the monoclinic space group C121, with unit-cell parameters a = 90.44, b = 44.52, c = 132.98 Å, β = 103.8°. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Kubiak, X. Pluvinage, B. Li de la Sierra-Gallay, I. Weber, P. Haouz, A. Dupret, J.-M. Rodrigues-Lima, F. 2012-01-26 doi:10.1107/S1744309111053942 International Union of Crystallography B. cereus arylamine N-acetyltransferase 3 was expressed, purified and crystallized. X-ray diffraction data were collected to 2.42 Å resolution and the crystals belonged to the monoclinic space group C121. EN arylamine N-acetyltransferases Bacillus cereus catalytic triad xenobiotic metabolism Arylamine N-acetyltransferases (NATs) are xenobiotic metabolizing enzymes (XMEs) that catalyze the acetylation of arylamines. All functional NATs described to date possess a strictly conserved Cys-His-Asp catalytic triad. Here, the purification, crystallization and preliminary X-ray characterization of Bacillus cereus arylamine N-acetyltransferase 3 [(BACCR)NAT3], a putative NAT isoenzyme that possesses a unique catalytic triad containing a glutamate residue, is reported. The crystal diffracted to 2.42 Å resolution and belonged to the monoclinic space group C121, with unit-cell parameters a = 90.44, b = 44.52, c = 132.98 Å, β = 103.8°. text/html Purification, crystallization and preliminary X-ray characterization of Bacillus cereus arylamine N-acetyltransferase 3 [(BACCR)NAT3] text 2 68 2012-01-26 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 196 198 http://scripts.iucr.org/cgi-bin/paper?xb5045 S-Ribosylhomocysteinase (LuxS) encoded by the luxS gene from Streptococcus mutans plays a crucial role in the quorum-sensing system. LuxS was solubly expressed in Escherichia coli with high yield. The purity of the purified target protein, which was identified by SDS–PAGE and MALDI–TOF MS analysis, was >95%. The protein was crystallized using the hanging-drop vapour-diffusion method with PEG 3350 as the primary precipitant. X-ray diffraction data were collected at Beijing Synchrotron Radiation Facility (BSRF). Diffraction by the crystal extended to 2.4 Å resolution and the crystal belonged to space group C2221, with unit-cell parameters a = 55.3, b = 148.7, c = 82.8 Å. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Li, H. Zhao, H. Zhu, L. Hong, L. Zhang, H. Lin, F. Xu, C. Li, S. Zhang, Z. 2012-01-26 doi:10.1107/S1744309111054212 International Union of Crystallography S-Ribosylhomocysteinase (LuxS) encoded by the LuxS gene from Streptococcus mutans was solubly expressed in Escherichia coli, purified and crystallized. Diffraction by the crystal extended to 2.4 Å resolution. EN Streptococcus mutans LuxS S-Ribosylhomocysteinase (LuxS) encoded by the luxS gene from Streptococcus mutans plays a crucial role in the quorum-sensing system. LuxS was solubly expressed in Escherichia coli with high yield. The purity of the purified target protein, which was identified by SDS–PAGE and MALDI–TOF MS analysis, was >95%. The protein was crystallized using the hanging-drop vapour-diffusion method with PEG 3350 as the primary precipitant. X-ray diffraction data were collected at Beijing Synchrotron Radiation Facility (BSRF). Diffraction by the crystal extended to 2.4 Å resolution and the crystal belonged to space group C2221, with unit-cell parameters a = 55.3, b = 148.7, c = 82.8 Å. text/html Crystallization and preliminary X-ray analysis of S-ribosylhomocysteinase from Streptococcus mutans text 2 68 2012-01-26 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 199 202 http://scripts.iucr.org/cgi-bin/paper?ft5017 Bacillus subtilis YwfE, an l-amino-acid ligase, catalyzes the formation of an α-dipeptide from l-amino acids in an ATP-dependent manner. In order to elucidate the substrate-recognition mode and the reaction mechanism of this ligase, native and selenomethionine-derivatized (SeMet) crystals of YwfE in the presence of ADP, MgCl2 and the dipeptide l-Ala-l-Gln were obtained using the hanging-drop vapour-diffusion method. These crystals diffracted to 1.9 and 2.8 Å resolution, respectively. Preliminary SAD phase calculations using the data set from the SeMet crystal suggested that the crystal belonged to the hexagonal space group P6522, with unit-cell parameters a = b = 90.85, c = 250.31 Å, and contained one molecule in the asymmetric unit with a solvent content of 57.3%. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Tsuda, T. Suzuki, T. Kojima, S. 2012-01-26 doi:10.1107/S174430911105425X International Union of Crystallography Native and selenomethionine-derivatized (SeMet) crystals of Bacillus subtilis YwfE in the presence of ADP, MgCl2 and the dipeptide l-Ala-l-Gln were obtained using the hanging-drop vapour-diffusion method. EN l-amino-acid ligases ATP-grasp fold Bacillus subtilis YwfE, an l-amino-acid ligase, catalyzes the formation of an α-dipeptide from l-amino acids in an ATP-dependent manner. In order to elucidate the substrate-recognition mode and the reaction mechanism of this ligase, native and selenomethionine-derivatized (SeMet) crystals of YwfE in the presence of ADP, MgCl2 and the dipeptide l-Ala-l-Gln were obtained using the hanging-drop vapour-diffusion method. These crystals diffracted to 1.9 and 2.8 Å resolution, respectively. Preliminary SAD phase calculations using the data set from the SeMet crystal suggested that the crystal belonged to the hexagonal space group P6522, with unit-cell parameters a = b = 90.85, c = 250.31 Å, and contained one molecule in the asymmetric unit with a solvent content of 57.3%. text/html Crystallization and preliminary X-ray diffraction analysis of Bacillus subtilis YwfE, an l-amino-acid ligase text 2 68 2012-01-26 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 203 206 http://scripts.iucr.org/cgi-bin/paper?fw5342 Aminopeptidases (APs) are a group of exopeptidases that catalyze the removal of amino acids from the N-termini of proteins and peptides. The APs are ubiquitous in nature and are of critical biological and medical importance because of their key role in protein degradation. Pseudomonas aeruginosa aspartyl aminopeptidase (PaAAP), which is encoded by the apeB gene, was expressed in Escherichia coli, purified and crystallized using the microbatch method. A preliminary structural study has been performed using the X-ray crystallographic method. The PaAAP crystal diffracted to 2.0 Å resolution and belonged to the rhombohedral space group H3, with unit-cell parameters a = b = 133.6, c = 321.2. The unit-cell volume of the crystal is compatible with the presence of four monomers in the asymmetric unit, with a corresponding Matthews coefficient VM of 2.95 Å3 Da−1 and a solvent content of 58.3%. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Natarajan, S. Mathews, R. 2012-01-26 doi:10.1107/S1744309111054388 International Union of Crystallography Aspartyl aminopeptidase (APP) is a metallopeptidase that catalyzes the hydrolysis of aspartic acid from the N-termini of polypeptide chains. The apeB gene from P. aeruginosa was cloned, the APP enzyme was expressed and crystallized and a preliminary X-ray crystallographic study was performed in order to understand its unique reaction pathway. EN Pseudomonas aeruginosa aspartyl aminopeptidase Aminopeptidases (APs) are a group of exopeptidases that catalyze the removal of amino acids from the N-termini of proteins and peptides. The APs are ubiquitous in nature and are of critical biological and medical importance because of their key role in protein degradation. Pseudomonas aeruginosa aspartyl aminopeptidase (PaAAP), which is encoded by the apeB gene, was expressed in Escherichia coli, purified and crystallized using the microbatch method. A preliminary structural study has been performed using the X-ray crystallographic method. The PaAAP crystal diffracted to 2.0 Å resolution and belonged to the rhombohedral space group H3, with unit-cell parameters a = b = 133.6, c = 321.2. The unit-cell volume of the crystal is compatible with the presence of four monomers in the asymmetric unit, with a corresponding Matthews coefficient VM of 2.95 Å3 Da−1 and a solvent content of 58.3%. text/html Cloning, expression, crystallization and preliminary X-ray crystallographic analysis of aspartyl aminopeptidase from the apeB gene of Pseudomonas aeruginosa text 2 68 2012-01-26 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 207 210 http://scripts.iucr.org/cgi-bin/paper?dp5017 Snake-venom l-amino-acid oxidases (SV-LAAOs) trigger a wide range of local and systematic effects, including inhibition of platelet aggregation, cytotoxicity, haemolysis, apoptosis and haemorrhage. These effects mainly arise from the uncontrolled release of the hydrogen peroxide that is produced by the redox reaction involving l-amino acids catalyzed by these flavoenzymes. Taking their clinical relevance into account, few SV-LAAOs have been structurally characterized and the structural determinants responsible for their broad direct and indirect pharmacological activities remain unclear. In this work, an LAAO from Bothrops jararacussu venom (BJu-LAAO) was purified and crystallized. The BJu-LAAO crystals belonged to space group P21, with unit-cell parameters a = 66.38, b = 72.19, c = 101.53 Å, β = 90.9°. The asymmetric unit contained two molecules and the structure was determined and partially refined at 3.0 Å resolution. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Ullah, A. Coronado, M. Murakami, M.T. Betzel, C. Arni, R.K. 2012-01-26 doi:10.1107/S1744309111054923 International Union of Crystallography An l-amino-acid oxidase from B. jararacussu venom was crystallized and X-ray diffraction data were collected to a maximum resolution of 3.0 Å. EN l-amino-acid oxidases snake venoms Snake-venom l-amino-acid oxidases (SV-LAAOs) trigger a wide range of local and systematic effects, including inhibition of platelet aggregation, cytotoxicity, haemolysis, apoptosis and haemorrhage. These effects mainly arise from the uncontrolled release of the hydrogen peroxide that is produced by the redox reaction involving l-amino acids catalyzed by these flavoenzymes. Taking their clinical relevance into account, few SV-LAAOs have been structurally characterized and the structural determinants responsible for their broad direct and indirect pharmacological activities remain unclear. In this work, an LAAO from Bothrops jararacussu venom (BJu-LAAO) was purified and crystallized. The BJu-LAAO crystals belonged to space group P21, with unit-cell parameters a = 66.38, b = 72.19, c = 101.53 Å, β = 90.9°. The asymmetric unit contained two molecules and the structure was determined and partially refined at 3.0 Å resolution. text/html Crystallization and preliminary X-ray diffraction analysis of an l-amino-acid oxidase from Bothrops jararacussu venom text 2 68 2012-01-26 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 211 213 http://scripts.iucr.org/cgi-bin/paper?bo5097 Dipeptidyl peptidase 10 (DPP10, DPPY) is an inactive peptidase associated with voltage-gated potassium channels, acting as a modulator of their electrophysiological properties, cell-surface expression and subcellular localization. Because potassium channels are important disease targets, biochemical and structural characterization of their interaction partners was sought. DPP10 was cloned and expressed using an insect-cell system and the protein was purified via His-tag affinity and size-exclusion chromatography. Crystals obtained by the sitting-drop method were orthorhombic, belonging to space group P212121 with unit-cell parameters a = 80.91, b = 143.73, c = 176.25 Å. A single solution with two molecules in the asymmetric unit was found using the structure of DPP6 (also called DPPX; PDB entry 1xfd) as the search model in a molecular replacement protocol. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Bezerra, G.A. Dobrovetsky, E. Seitova, A. Dhe-Paganon, S. Gruber, K. 2012-01-26 doi:10.1107/S1744309111055230 International Union of Crystallography The expression, purification and crystallization of human dipeptidyl peptidase 10, a component of voltage-gated potassium channels, is described. EN dipeptidyl peptidase 10 voltage-gated potassium channels Dipeptidyl peptidase 10 (DPP10, DPPY) is an inactive peptidase associated with voltage-gated potassium channels, acting as a modulator of their electrophysiological properties, cell-surface expression and subcellular localization. Because potassium channels are important disease targets, biochemical and structural characterization of their interaction partners was sought. DPP10 was cloned and expressed using an insect-cell system and the protein was purified via His-tag affinity and size-exclusion chromatography. Crystals obtained by the sitting-drop method were orthorhombic, belonging to space group P212121 with unit-cell parameters a = 80.91, b = 143.73, c = 176.25 Å. A single solution with two molecules in the asymmetric unit was found using the structure of DPP6 (also called DPPX; PDB entry 1xfd) as the search model in a molecular replacement protocol. text/html Crystallization and preliminary X-ray diffraction analysis of human dipeptidyl peptidase 10 (DPPY), a component of voltage-gated potassium channels text 2 68 2012-01-26 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 214 217 http://scripts.iucr.org/cgi-bin/paper?nj5107 Deinococcus radiodurans has developed an efficient mechanism which allows the integrity of its entire genome to be fully restored after exposure to very high doses of ionizing radiation. Homologous recombination plays a crucial role in this process. RecN is a protein that belongs to the SMC-like protein family and is suggested to be involved in DNA repair. RecN is composed of a globular domain and an antiparallel coiled-coil region which connects the N- and C-termini. It has been suggested that dimerization of RecN occurs via the coiled-coil domain, but to date there is no structural or biochemical evidence for this. Here, SAXS studies and preliminary X-ray diffraction data of crystals of the purified coiled-coil domain of RecN are presented. The structure was solved by single-wavelength anomalous dispersion using SeMet derivatives, and preliminary electron-density maps support the rod-like model derived from the SAXS data. Model building and refinement are still ongoing. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Pellegrino, S. de Sanctis, D. McSweeney, S. Timmins, J. 2012-01-26 doi:10.1107/S1744309111055187 International Union of Crystallography The coiled-coil domain of the essential DNA-repair protein RecN from D. radiodurans was crystallized. The crystals belonged to space group P21 and diffracted X-rays to 2.04 Å resolution. EN DNA repair homologous recombination RecN SMC-like proteins Deinococcus radiodurans has developed an efficient mechanism which allows the integrity of its entire genome to be fully restored after exposure to very high doses of ionizing radiation. Homologous recombination plays a crucial role in this process. RecN is a protein that belongs to the SMC-like protein family and is suggested to be involved in DNA repair. RecN is composed of a globular domain and an antiparallel coiled-coil region which connects the N- and C-termini. It has been suggested that dimerization of RecN occurs via the coiled-coil domain, but to date there is no structural or biochemical evidence for this. Here, SAXS studies and preliminary X-ray diffraction data of crystals of the purified coiled-coil domain of RecN are presented. The structure was solved by single-wavelength anomalous dispersion using SeMet derivatives, and preliminary electron-density maps support the rod-like model derived from the SAXS data. Model building and refinement are still ongoing. text/html Expression, purification and preliminary structural analysis of the coiled-coil domain of Deinococcus radiodurans RecN text 2 68 2012-01-26 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 218 221 http://scripts.iucr.org/cgi-bin/paper?pu5354 Protein E (PE) is a ubiquitous multifunctional surface protein of Haemophilus spp. and other bacterial pathogens of the Pasteurellaceae family. H. influenzae utilizes PE for attachment to respiratory epithelial cells. In addition, PE interacts directly with plasminogen and the extracellular matrix (ECM) components vitronectin and laminin. Vitronectin is a complement regulator that inhibits the formation of the membrane-attack complex (MAC). PE-mediated vitronectin recruitment at the H. influenzae surface thus inhibits MAC and protects against serum bactericidal activity. Laminin is an abundant ECM protein and is present in the basement membrane that helps in adherence of H. influenzae during colonization. Here, the expression, purification and crystallization of and the collection of high-resolution data for this important H. influenzae adhesin are reported. To solve the phase problem for PE, Met residues were introduced and an SeMet variant was expressed and crystallized. Both native and SeMet-containing PE gave plate-like crystals in space group P21, with unit-cell parameters a = 44, b = 57, c = 61 Å, β = 96°. Diffraction data collected from native and SeMet-derivative crystals extended to resolutions of 1.8 and 2.6 Å, respectively. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Singh, B. Al Jubair, T. Förnvik, K. Thunnissen, M.M. Riesbeck, K. 2012-01-27 doi:10.1107/S1744309111055503 International Union of Crystallography Protein E of the respiratory pathogen H. influenzae is a multifunctional adhesin that is involved in bacterial attachment to host epithelium and direct interactions with vitronectin, laminin and plasminogen. The method of crystallization and X-ray data collection for protein E at 1.8 Å is presented. EN protein E Haemophilus influenzae surface-adhesion proteins Protein E (PE) is a ubiquitous multifunctional surface protein of Haemophilus spp. and other bacterial pathogens of the Pasteurellaceae family. H. influenzae utilizes PE for attachment to respiratory epithelial cells. In addition, PE interacts directly with plasminogen and the extracellular matrix (ECM) components vitronectin and laminin. Vitronectin is a complement regulator that inhibits the formation of the membrane-attack complex (MAC). PE-mediated vitronectin recruitment at the H. influenzae surface thus inhibits MAC and protects against serum bactericidal activity. Laminin is an abundant ECM protein and is present in the basement membrane that helps in adherence of H. influenzae during colonization. Here, the expression, purification and crystallization of and the collection of high-resolution data for this important H. influenzae adhesin are reported. To solve the phase problem for PE, Met residues were introduced and an SeMet variant was expressed and crystallized. Both native and SeMet-containing PE gave plate-like crystals in space group P21, with unit-cell parameters a = 44, b = 57, c = 61 Å, β = 96°. Diffraction data collected from native and SeMet-derivative crystals extended to resolutions of 1.8 and 2.6 Å, respectively. text/html Crystallization and X-ray diffraction analysis of a novel surface-adhesin protein: protein E from Haemophilus influenzae text 2 68 2012-01-27 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 222 226 http://scripts.iucr.org/cgi-bin/paper?dp5016 Clostridium botulinum produces botulinum neurotoxin (BoNT) as a large toxin complex assembled with nontoxic nonhaemagglutinin (NTNHA) and/or haemagglutinin components. Complex formation with NTNHA is considered to be critical in eliciting food poisoning because the complex shields the BoNT from the harsh conditions in the digestive tract. In the present study, NTNHA was expressed in Escherichia coli and crystallized. Diffraction data were collected to 3.9 Å resolution. The crystal belonged to the trigonal space group P321 or P3121/P3221, with unit-cell parameters a = b = 147.85, c = 229.74 Å. The structure of NTNHA will provide insight into the assembly mechanism that produces the unique BoNT–NTNHA complex. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Miyata, K. Inui, K. Miyashita, S.-I. Sagane, Y. Hasegawa, K. Matsumoto, T. Yamano, A. Niwa, K. Watanabe, T. Ohyama, T. 2012-01-27 doi:10.1107/S174430911105603X International Union of Crystallography The nontoxic nonhaemagglutinin of serotype D neurotoxin complex, which is important in proteolytic stability for toxin complex, was expressed in E. coli, and crystallized. EN Clostridium botulinum nontoxic nonhaemagglutinin botulinum toxin complexes spontaneous nicking Clostridium botulinum produces botulinum neurotoxin (BoNT) as a large toxin complex assembled with nontoxic nonhaemagglutinin (NTNHA) and/or haemagglutinin components. Complex formation with NTNHA is considered to be critical in eliciting food poisoning because the complex shields the BoNT from the harsh conditions in the digestive tract. In the present study, NTNHA was expressed in Escherichia coli and crystallized. Diffraction data were collected to 3.9 Å resolution. The crystal belonged to the trigonal space group P321 or P3121/P3221, with unit-cell parameters a = b = 147.85, c = 229.74 Å. The structure of NTNHA will provide insight into the assembly mechanism that produces the unique BoNT–NTNHA complex. text/html Crystallization and preliminary X-ray analysis of the Clostridium botulinum type D nontoxic nonhaemagglutinin text 2 68 2012-01-27 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 227 230 http://scripts.iucr.org/cgi-bin/paper?rl5016 FrpB is an integral outer membrane protein from the human pathogen Neisseria meningitidis. It is a member of the TonB-dependent transporter family and promotes the uptake of iron across the outer membrane. There is also evidence that FrpB is an antigen and hence a potential component of a vaccine against meningococcal meningitis. FrpB incorporating a polyhistidine tag was overexpressed in Escherichia coli into inclusion bodies. The protein was then solubilized in urea, refolded and purified to homogeneity. Two separate antigenic variants of FrpB were crystallized by sitting-drop vapour diffusion. Crystals of the F5-1 variant diffracted to 2.4 Å resolution and belonged to space group C2, with unit-cell parameters a = 176.5, b = 79.4, c = 75.9 Å, β = 98.3°. Crystal-packing calculations suggested the presence of a monomer in the asymmetric unit. Crystals of the F3-3 variant also diffracted to 2.4 Å resolution and belonged to space group P212121, with unit-cell parameters a = 85.3, b = 104.6, c = 269.1 Å. Preliminary analysis suggested the presence of an FrpB trimer in the asymmetric unit. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Saleem, M. Prince, S.M. Patel, H. Chan, H. Feavers, I.M. Derrick, J.P. 2012-01-27 doi:10.1107/S1744309111056028 International Union of Crystallography The refolding, purification and crystallization of FrpB from the meningitis pathogen Neisseria meningitidis is described. EN outer membrane proteins FrpB Neisseria meningitidis FrpB is an integral outer membrane protein from the human pathogen Neisseria meningitidis. It is a member of the TonB-dependent transporter family and promotes the uptake of iron across the outer membrane. There is also evidence that FrpB is an antigen and hence a potential component of a vaccine against meningococcal meningitis. FrpB incorporating a polyhistidine tag was overexpressed in Escherichia coli into inclusion bodies. The protein was then solubilized in urea, refolded and purified to homogeneity. Two separate antigenic variants of FrpB were crystallized by sitting-drop vapour diffusion. Crystals of the F5-1 variant diffracted to 2.4 Å resolution and belonged to space group C2, with unit-cell parameters a = 176.5, b = 79.4, c = 75.9 Å, β = 98.3°. Crystal-packing calculations suggested the presence of a monomer in the asymmetric unit. Crystals of the F3-3 variant also diffracted to 2.4 Å resolution and belonged to space group P212121, with unit-cell parameters a = 85.3, b = 104.6, c = 269.1 Å. Preliminary analysis suggested the presence of an FrpB trimer in the asymmetric unit. text/html Refolding, purification and crystallization of the FrpB outer membrane iron transporter from Neisseria meningitidis text 2 68 2012-01-27 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 231 235 http://scripts.iucr.org/cgi-bin/paper?en5486 Chaperones promote many different molecular processes, including the folding, targeting and degradation of proteins. The best-studied chaperone system consists of the Hsp70s and their co-chaperones the Hsp40s. Chaperone function can be hijacked by viruses in plants. Potato virus Y interacts via its coat protein with an Hsp40 from Nicotiana tabacum, referred to as NtCPIP1, in order to regulate replication. To understand the molecular determinants of this mechanism, different variants of NtCPIP1 were expressed, purified and crystallized. While crystals of wild-type NtCPIP1 diffracted to 8.0 Å resolution, the deletion mutant NtCPIP1-Δ(1:127) crystallized in space group P21212 and diffracted to 2.4 Å resolution. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Griessl, M.H. Jungkunz, I. Sonnewald, U. Muller, Y.A. 2012-01-27 doi:10.1107/S1744309111055928 International Union of Crystallography CPIP1 from N. tabacum (NtCPIP1) is a plant heat-shock 40-type protein that participates in effector–host interactions during potyviral infections. Whereas crystals of full-length wild-type NtCPIP1 only diffracted to 8.0 Å resolution, diffraction from crystals of a shortened variant lacking the first 127 amino acids extended to 2.4 Å resolution. EN plant heat-shock protein 40 viral infections potyviruses Chaperones promote many different molecular processes, including the folding, targeting and degradation of proteins. The best-studied chaperone system consists of the Hsp70s and their co-chaperones the Hsp40s. Chaperone function can be hijacked by viruses in plants. Potato virus Y interacts via its coat protein with an Hsp40 from Nicotiana tabacum, referred to as NtCPIP1, in order to regulate replication. To understand the molecular determinants of this mechanism, different variants of NtCPIP1 were expressed, purified and crystallized. While crystals of wild-type NtCPIP1 diffracted to 8.0 Å resolution, the deletion mutant NtCPIP1-Δ(1:127) crystallized in space group P21212 and diffracted to 2.4 Å resolution. text/html Purification, crystallization and preliminary X-ray diffraction analysis of the Hsp40 protein CPIP1 from Nicotiana tabacum text 2 68 2012-01-27 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 236 239 http://scripts.iucr.org/cgi-bin/paper?fw5335 A ribokinase gene (rbk) from the anaerobic halothermophilic bacterium Halothermothrix orenii was cloned and overexpressed in Escherichia coli. The recombinant protein (Ho-Rbk) was purified using immobilized metal-ion affinity chromatography and crystals were obtained using the sitting-drop method. Diffraction data were collected to a resolution of 3.1 Å using synchrotron radiation. The crystals belonged to the orthorhombic space group P212121, with unit-cell parameters a = 45.6, b = 61.1, c = 220.2, and contained two molecules per asymmetric unit. A molecular-replacement solution has been found and attempts are currently under way to build a model of the ribokinase. Efforts to improve crystal quality so that higher resolution data can be obtained are also being considered. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Kori, L.D. Hofmann, A. Patel, B.K.C. 2012-01-27 doi:10.1107/S1744309111041091 International Union of Crystallography A ribokinase gene (rbk) from the anaerobic halothermophilic bacterium Halothermothrix orenii was cloned and overexpressed in Escherichia coli. The recombinant protein (Ho-Rbk) was purified using immobilized metal-ion affinity chromatography and crystals were obtained using the sitting-drop method. EN ribokinases sugar kinases halothermophiles Halothermothrix orenii A ribokinase gene (rbk) from the anaerobic halothermophilic bacterium Halothermothrix orenii was cloned and overexpressed in Escherichia coli. The recombinant protein (Ho-Rbk) was purified using immobilized metal-ion affinity chromatography and crystals were obtained using the sitting-drop method. Diffraction data were collected to a resolution of 3.1 Å using synchrotron radiation. The crystals belonged to the orthorhombic space group P212121, with unit-cell parameters a = 45.6, b = 61.1, c = 220.2, and contained two molecules per asymmetric unit. A molecular-replacement solution has been found and attempts are currently under way to build a model of the ribokinase. Efforts to improve crystal quality so that higher resolution data can be obtained are also being considered. text/html Expression, purification, crystallization and preliminary X-ray diffraction analysis of a ribokinase from the thermohalophile Halothermothrix orenii text 2 68 2012-01-27 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 240 243 http://scripts.iucr.org/cgi-bin/paper?fw5336 Orotate phosphoribosyltransferase (OPRT) catalyzes the Mg2+-dependent condensation of orotic acid (OA) with 5-α-d-phosphorylribose 1-diphosphate (PRPP) to yield diphosphate (PPi) and the nucleotide orotidine 5′-monophosphate. OPRT from Plasmodium falciparum produced in Escherichia coli was crystallized by the sitting-drop vapour-diffusion method in complex with OA and PRPP in the presence of Mg2+. The crystal exhibited tetragonal symmetry, belonging to space group P41 or P43, with unit-cell parameters a = b = 49.15, c = 226.94 Å. X-ray diffraction data were collected to 2.5 Å resolution at 100 K using a synchrotron-radiation source. Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 Takashima, Y. Mizohata, E. Tokuoka, K. Krungkrai, S.R. Kusakari, Y. Konishi, S. Satoh, A. Matsumura, H. Krungkrai, J. Horii, T. Inoue, T. 2012-01-27 doi:10.1107/S1744309111043247 International Union of Crystallography Orotate phosphoribosyltransferase from Plasmodium falciparum produced in Escherichia coli was crystallized by the sitting-drop vapour-diffusion method in complex with OA and PRPP in the presence of Mg2+. EN orotate phosphoribosyltransferase Plasmodium falciparum Orotate phosphoribosyltransferase (OPRT) catalyzes the Mg2+-dependent condensation of orotic acid (OA) with 5-α-d-phosphorylribose 1-diphosphate (PRPP) to yield diphosphate (PPi) and the nucleotide orotidine 5′-monophosphate. OPRT from Plasmodium falciparum produced in Escherichia coli was crystallized by the sitting-drop vapour-diffusion method in complex with OA and PRPP in the presence of Mg2+. The crystal exhibited tetragonal symmetry, belonging to space group P41 or P43, with unit-cell parameters a = b = 49.15, c = 226.94 Å. X-ray diffraction data were collected to 2.5 Å resolution at 100 K using a synchrotron-radiation source. text/html Crystallization and preliminary X-ray diffraction analysis of orotate phosphoribosyltransferase from the human malaria parasite Plasmodium falciparum text 2 68 2012-01-27 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 244 246 http://scripts.iucr.org/cgi-bin/paper?me0457 Copyright (c) 2012 International Union of Crystallography urn:issn:1744-3091 IUCr Editorial Office 2012-01-28 doi:10.1107/S1744309111042448 International Union of Crystallography Notes for authors. EN Notes for authors text/html Notes for authors 2012 text 2 68 2012-01-28 Copyright (c) 2012 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications international union of crystallography 247 251