http://journals.iucr.org/f/issues/2009/07/00/isscontsbdy.html Acta Crystallographica Section F Structural Biology and Crystallization Communications aims to provide a home for communications on the crystallization and structure determination of biological macromolecules. It will commence publication in January 2005 and will include three categories of publication: Structural genomics communications, Protein structure communications, Crystallization communications. Articles will be available online when ready, making publication as fast as possible, and will include unlimited free colour illustrations, movies and other enhancements. The editorial process will be completely electronic with respect to deposition, submission, refereeing and publication. en Copyright (c) 2009 International Union of Crystallography 2009-07-01 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 aims to provide a home for communications on the crystallization and structure determination of biological macromolecules. It will commence publication in January 2005 and will include three categories of publication: Structural genomics communications, Protein structure communications, Crystallization communications. Articles will be available online when ready, making publication as fast as possible, and will include unlimited free colour illustrations, movies and other enhancements. The editorial process will be completely electronic with respect to deposition, submission, refereeing and publication. text/html Acta Crystallographica Section F: Structural Biology and Crystallization Communications, Volume 65, Part 7, 2009 text monthly 1 2002-01-01T00:00+00:00 7 65 2009-07-01 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications 654 urn:issn:1744-3091 med@iucr.org July 2009 2009-07-01 http://journals.iucr.org/logos/rss10f.gif http://journals.iucr.org/f/issues/2009/07/00/isscontsbdy.html Still image http://scripts.iucr.org/cgi-bin/paper?fw5214 A primary role for mitochondrial dysfunction is indicated in the pathogenesis of insulin resistance. A widely used drug for the treatment of type 2 diabetes is pioglitazone, a member of the thiazolidinedione class of molecules. MitoNEET, a 2Fe–2S outer mitochondrial membrane protein, binds pioglitazone [Colca et al. (2004), Am. J. Physiol. Endocrinol. Metab. 286, E252–E260]. The soluble domain of the human mitoNEET protein has been expressed C-terminal to the superfolder green fluorescent protein and the mitoNEET protein has been isolated. Comparison of the crystal structure of mitoNEET isolated from cleavage of the fusion protein (1.4 Å resolution, R factor = 20.2%) with other solved structures shows that the CDGSH domains are superimposable, indicating proper assembly of mitoNEET. Furthermore, there is considerable flexibility in the position of the cytoplasmic tethering arms, resulting in two different conformations in the crystal structure. This flexibility affords multiple orientations on the outer mitochondrial membrane. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Conlan, A.R. Paddock, M.L. Axelrod, H.L. Cohen, A.E. Abresch, E.C. Wiley, S. Roy, M. Nechushtai, R. Jennings, P.A. 2009-06-27 doi:10.1107/S1744309109019605 International Union of Crystallography The crystal structure of the anti-diabetic drug target mitoNEET obtained from a GFP fusion construct (1.4 Å resolution, R factor = 20.2%) shows that the CDGSH 2Fe–2S binding domains are superimposable with previously determined non-fused constructs. However, there is considerable flexibility in the position of the outer mitochondrial tethering arms resulting in two different conformations in the crystal structure. en mitochondrial outer membrane 2Fe–2S proteins anti-diabetic drug target CDGSH domain expression A primary role for mitochondrial dysfunction is indicated in the pathogenesis of insulin resistance. A widely used drug for the treatment of type 2 diabetes is pioglitazone, a member of the thiazolidinedione class of molecules. MitoNEET, a 2Fe–2S outer mitochondrial membrane protein, binds pioglitazone [Colca et al. (2004), Am. J. Physiol. Endocrinol. Metab. 286, E252–E260]. The soluble domain of the human mitoNEET protein has been expressed C-terminal to the superfolder green fluorescent protein and the mitoNEET protein has been isolated. Comparison of the crystal structure of mitoNEET isolated from cleavage of the fusion protein (1.4 Å resolution, R factor = 20.2%) with other solved structures shows that the CDGSH domains are superimposable, indicating proper assembly of mitoNEET. Furthermore, there is considerable flexibility in the position of the cytoplasmic tethering arms, resulting in two different conformations in the crystal structure. This flexibility affords multiple orientations on the outer mitochondrial membrane. text/html The novel 2Fe–2S outer mitochondrial protein mitoNEET displays conformational flexibility in its N-terminal cytoplasmic tethering domain text 7 65 2009-06-27 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications structural communications 654 659 http://scripts.iucr.org/cgi-bin/paper?dz5163 The crystal structure of an echinomycin–d(ACGTACGT) duplex interacting with manganese(II) was solved by Mn-SAD using in-house data and refined to 1.1 Å resolution against synchrotron data. This complex crystallizes in a different space group compared with related complexes and shows a different mode of base pairing next to the bis-intercalation site, suggesting that the energy difference between Hoogsteen and Watson–Crick pairing is rather small. The binding of manganese to N7 of guanine is only possible because of DNA unwinding induced by the echinomycin, which might help to explain the mode of action of the drug. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Pfoh, R. Cuesta-Seijo, J.A. Sheldrick, G.M. 2009-06-27 doi:10.1107/S1744309109019654 International Union of Crystallography In the presence of Mn2+, a new crystal form of an echinomycin–d(ACGTACGT) complex is found which shows mixed base pairing next to the bis-intercalation site. en echinomycin manganese SAD phasing DNA unwinding The crystal structure of an echinomycin–d(ACGTACGT) duplex interacting with manganese(II) was solved by Mn-SAD using in-house data and refined to 1.1 Å resolution against synchrotron data. This complex crystallizes in a different space group compared with related complexes and shows a different mode of base pairing next to the bis-intercalation site, suggesting that the energy difference between Hoogsteen and Watson–Crick pairing is rather small. The binding of manganese to N7 of guanine is only possible because of DNA unwinding induced by the echinomycin, which might help to explain the mode of action of the drug. text/html Interaction of an echinomycin–DNA complex with manganese ions text 7 65 2009-06-27 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications structural communications 660 664 http://scripts.iucr.org/cgi-bin/paper?hv5132 Proteins of the A-type lamin family, which consists of two members, lamin A and lamin C, are the major components of a thin proteinaceous filamentous meshwork, the lamina, that underlies the inner nuclear membrane. A-type lamins have recently become the focus of extensive functional studies as a consequence of the linking of at least eight congenital diseases to mutations in the lamin A/C gene (LMNA). This spectrum of pathologies, which mostly manifest themselves as dominant traits, includes muscle dystrophies, dilated cardiomyopathies, the premature aging syndrome Hutchinson–Guilford progeria and familial partial lipodystrophy (FPLD). The crystal structure of the lamin A/C mutant R482W, a variant that causes FPLD, has been determined at 1.5 Å resolution. A completely novel aggregation state of the C-terminal globular domain and the position of the mutated amino-acid residue suggest means by which the mutation may affect lamin A/C–protein and protein–DNA interactions. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Magracheva, E. Kozlov, S. Stewart, C.L. Wlodawer, A. Zdanov, A. 2009-06-27 doi:10.1107/S1744309109020302 International Union of Crystallography The crystal structure of the R482W mutant of the C-terminal domain of lamin A/C, which is responsible for familial partial lipodystrophy, has been determined at 1.5 Å resolution. A completely novel aggregation state of the mutated protein may be responsible for the changes in biological activity. en lamin A/C familial partial lipodystrophy Proteins of the A-type lamin family, which consists of two members, lamin A and lamin C, are the major components of a thin proteinaceous filamentous meshwork, the lamina, that underlies the inner nuclear membrane. A-type lamins have recently become the focus of extensive functional studies as a consequence of the linking of at least eight congenital diseases to mutations in the lamin A/C gene (LMNA). This spectrum of pathologies, which mostly manifest themselves as dominant traits, includes muscle dystrophies, dilated cardiomyopathies, the premature aging syndrome Hutchinson–Guilford progeria and familial partial lipodystrophy (FPLD). The crystal structure of the lamin A/C mutant R482W, a variant that causes FPLD, has been determined at 1.5 Å resolution. A completely novel aggregation state of the C-terminal globular domain and the position of the mutated amino-acid residue suggest means by which the mutation may affect lamin A/C–protein and protein–DNA interactions. text/html Structure of the lamin A/C R482W mutant responsible for dominant familial partial lipodystrophy (FPLD) text 7 65 2009-06-27 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications structural communications 665 670 http://scripts.iucr.org/cgi-bin/paper?en5366 In the quest for the rational design of selective and potent inhibitors for members of the pancreatic ribonuclease A (RNase A) family of biomedical interest, the binding of uridine 5′-phosphate (U5P) and uridine 5′-diphosphate (UDP) to RNase A have been investigated using kinetic studies and X-ray crystallography. Both nucleotides are competitive inhibitors of the enzyme, with Ki values of 4.0 and 0.65 mM, respectively. They bind to the active site of the enzyme by anchoring two molecules connected to each other by hydrogen bonds and van der Waals interactions. While the first of the inhibitor molecules binds with its nucleobase in the pyrimidinyl-binding subsite, the second is bound at the purine-preferring subsite. The unexpected binding of a pyrimidine at the purine-binding subsite has added new important elements to the rational design approach for the discovery of new potent inhibitors of the RNase A superfamily. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Tsirkone, V.G. Dossi, K. Drakou, C. Zographos, S.E. Kontou, M. Leonidas, D.D. 2009-06-27 doi:10.1107/S1744309109021423 International Union of Crystallography The structure of ribonuclease A in complex with uridine 5′-phosphate and uridine 5′-diphosphate has been determined at 1.4 Å resolution in order to facilitate the rational design of selective and potent inhibitors. en ribnuclease A uridine 5′-phosphate uridine 5′-diphosphate inhibitor design In the quest for the rational design of selective and potent inhibitors for members of the pancreatic ribonuclease A (RNase A) family of biomedical interest, the binding of uridine 5′-phosphate (U5P) and uridine 5′-diphosphate (UDP) to RNase A have been investigated using kinetic studies and X-ray crystallography. Both nucleotides are competitive inhibitors of the enzyme, with Ki values of 4.0 and 0.65 mM, respectively. They bind to the active site of the enzyme by anchoring two molecules connected to each other by hydrogen bonds and van der Waals interactions. While the first of the inhibitor molecules binds with its nucleobase in the pyrimidinyl-binding subsite, the second is bound at the purine-preferring subsite. The unexpected binding of a pyrimidine at the purine-binding subsite has added new important elements to the rational design approach for the discovery of new potent inhibitors of the RNase A superfamily. text/html Inhibitor design for ribonuclease A: the binding of two 5′-phosphate uridine analogues text 7 65 2009-06-27 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications structural communications 671 677 http://scripts.iucr.org/cgi-bin/paper?bw5294 γ-Glutamylcysteine synthetase–glutathione synthetase (γGCS-GS) is a bifunctional enzyme that catalyzes two consecutive steps of ATP-dependent peptide formation in glutathione biosynthesis. Streptococcus agalactiae γGCS-GS is a target for the development of potential therapeutic agents. γGCS-GS was crystallized using the sitting-drop vapour-diffusion method. The crystals grew to dimensions of 0.3 × 0.2 × 0.2 mm under reducing conditions with 5 mM TCEP. X-ray data were collected to 2.8 Å resolution from a tetragonal crystal that belonged to space group I41. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Nakashima, Y. Nii, H. Janowiak, B.E. Griffith, O.W. Hibi, T. 2009-06-27 doi:10.1107/S1744309109018636 International Union of Crystallography The S. agalactiae bifunctional ligase for glutathione synthesis has been crystallized. Native data were collected to 2.8 Å resolution. en Gram-positive bacteria glutathione synthesis chemotherapy γ-Glutamylcysteine synthetase–glutathione synthetase (γGCS-GS) is a bifunctional enzyme that catalyzes two consecutive steps of ATP-dependent peptide formation in glutathione biosynthesis. Streptococcus agalactiae γGCS-GS is a target for the development of potential therapeutic agents. γGCS-GS was crystallized using the sitting-drop vapour-diffusion method. The crystals grew to dimensions of 0.3 × 0.2 × 0.2 mm under reducing conditions with 5 mM TCEP. X-ray data were collected to 2.8 Å resolution from a tetragonal crystal that belonged to space group I41. text/html Crystallization and preliminary crystallographic analysis of bifunctional γ-glutamylcysteine synthetase–glutatione synthetase from Streptococcus agalactiae text 7 65 2009-06-27 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 678 680 http://scripts.iucr.org/cgi-bin/paper?pu5261 Haemoglobin is a tetrameric protein that carries oxygen from the lungs to tissues and carbon dioxide from tissues back to the lungs. The oxygen-binding properties of haemoglobin are regulated through the binding of allosteric effectors. The respiratory system of avian species is unique and complex in nature when compared with that of mammals. In avian species, inositol pentaphosphate (inositol-P5) is present in the erythrocytes of the adult and is thought to be the major factor responsible for the relatively high oxygen affinity of the whole blood. The ostrich (Struthio camelus) is a large flightless bird which contains inositol tetrakisphosphate (inositol-P4) in its erythrocytes and its whole blood oxygen affinity is higher. Efforts have been made to explore the structure–function relationship of ostrich haemoglobin. Ostrich haemoglobin was purified using ion-exchange chromatography. Haemoglobin crystals were grown by the hanging-drop vapour-diffusion method using PEG 3350 as the precipitant in 50 mM phosphate buffer pH 7.2. Data were collected using a MAR345 image-plate detector system. The crystals of ostrich haemoglobin diffracted to 2.2 Å resolution. They belonged to the orthorhombic space group P212121 with one whole biological molecule in the asymmetric unit; the unit-cell parameters were a = 80.93, b = 81.68, c = 102.05 Å. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Sundaresan, S.S. Ramesh, P. Sivakumar, K. Ponnuswamy, M.N. 2009-06-27 doi:10.1107/S1744309109019009 International Union of Crystallography Purification, crystallization and preliminary X-ray analysis of haemoglobin from ostrich (Struthio camelus) has been carried out under 293 K temperature conditions. The ostrich is a large flightless bird which contains inositol tetrakisphosphate in erythrocytes and its whole blood oxygen affinity is higher. Efforts have been made to explore the structure–function relationship of ostrich heamoglobin. en haemoglobin Struthio camelus Haemoglobin is a tetrameric protein that carries oxygen from the lungs to tissues and carbon dioxide from tissues back to the lungs. The oxygen-binding properties of haemoglobin are regulated through the binding of allosteric effectors. The respiratory system of avian species is unique and complex in nature when compared with that of mammals. In avian species, inositol pentaphosphate (inositol-P5) is present in the erythrocytes of the adult and is thought to be the major factor responsible for the relatively high oxygen affinity of the whole blood. The ostrich (Struthio camelus) is a large flightless bird which contains inositol tetrakisphosphate (inositol-P4) in its erythrocytes and its whole blood oxygen affinity is higher. Efforts have been made to explore the structure–function relationship of ostrich haemoglobin. Ostrich haemoglobin was purified using ion-exchange chromatography. Haemoglobin crystals were grown by the hanging-drop vapour-diffusion method using PEG 3350 as the precipitant in 50 mM phosphate buffer pH 7.2. Data were collected using a MAR345 image-plate detector system. The crystals of ostrich haemoglobin diffracted to 2.2 Å resolution. They belonged to the orthorhombic space group P212121 with one whole biological molecule in the asymmetric unit; the unit-cell parameters were a = 80.93, b = 81.68, c = 102.05 Å. text/html Purification, crystallization and preliminary X-ray analysis of haemoglobin from ostrich (Struthio camelus) text 7 65 2009-06-27 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 681 683 http://scripts.iucr.org/cgi-bin/paper?rp5037 The transmembrane protein FeoB plays a key role in ferrous iron acquisition in prokaryotes. The N-terminal domain of FeoB from Methanococcus jannaschii was overproduced, purified to homogeneity and crystallized in the presence of GTP and magnesium. The native protein crystallized in a tetragonal space group and the crystals diffracted to beyond 2.2 Å resolution, with unit-cell parameters a = b = 84.77, c = 137.90 Å. The Matthews coefficient and the solvent content were estimated to be 2.65 Å3 Da−1 and 53.64%, respectively, which corresponds to the presence of two molecules per asymmetric unit. To obtain initial phases, selenomethionyl-substituted protein was overproduced, purified and crystallized. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Köster, S. Kühlbrandt, W. Yildiz, Ö. 2009-06-27 doi:10.1107/S1744309109019216 International Union of Crystallography The N-terminal domain of FeoB from Methanococcus jannaschii was overproduced, purified to homogeneity and crystallized in the presence of GTP and magnesium. en FeoB G domains Methanococcus jannaschii membrane proteins The transmembrane protein FeoB plays a key role in ferrous iron acquisition in prokaryotes. The N-terminal domain of FeoB from Methanococcus jannaschii was overproduced, purified to homogeneity and crystallized in the presence of GTP and magnesium. The native protein crystallized in a tetragonal space group and the crystals diffracted to beyond 2.2 Å resolution, with unit-cell parameters a = b = 84.77, c = 137.90 Å. The Matthews coefficient and the solvent content were estimated to be 2.65 Å3 Da−1 and 53.64%, respectively, which corresponds to the presence of two molecules per asymmetric unit. To obtain initial phases, selenomethionyl-substituted protein was overproduced, purified and crystallized. text/html Purification, crystallization and preliminary X-ray diffraction analysis of the FeoB G domain from Methanococcus jannaschii text 7 65 2009-06-27 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 684 687 http://scripts.iucr.org/cgi-bin/paper?bw5293 The tetramerization domain (T1 domain) derived from the voltage-dependent potassium channel Kv1.3 of Homo sapiens was recombinantly expressed in Escherichia coli and purified. The crystals were first grown in an NMR tube in 150 mM potassium phosphate pH 6.5 in the absence of additional precipitants. The crystals showed I4 symmetry characteristic of the naturally occurring tetrameric assembly of the single subunits. A complete native data set was collected to 1.2 Å resolution at 100 K using synchrotron radiation. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Winklmeier, A. Weyand, M. Schreier, C. Kalbitzer, H.R. Kremer, W. 2009-06-27 doi:10.1107/S1744309109019514 International Union of Crystallography The tetramerization domain of human Kv1.3 was cloned, expressed, purified and crystallized. The crystals belonged to space group I4 and diffracted to 1.2 Å resolution using synchrotron radiation. en human Kv1.3 potassium channels tetramerization domain The tetramerization domain (T1 domain) derived from the voltage-dependent potassium channel Kv1.3 of Homo sapiens was recombinantly expressed in Escherichia coli and purified. The crystals were first grown in an NMR tube in 150 mM potassium phosphate pH 6.5 in the absence of additional precipitants. The crystals showed I4 symmetry characteristic of the naturally occurring tetrameric assembly of the single subunits. A complete native data set was collected to 1.2 Å resolution at 100 K using synchrotron radiation. text/html Crystallization and preliminary X-ray diffraction studies of the tetramerization domain derived from the human potassium channel Kv1.3 text 7 65 2009-06-27 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 688 691 http://scripts.iucr.org/cgi-bin/paper?hc5083 ErbB2 is a transmembrane tyrosine kinase, the overexpression of which causes abnormality and disorder in cell signalling and leads to cell transformation. Previously, an anti-ErbB2 single-chain chimeric antibody chA21 that specifically inhibits the growth of ErbB2-overexpressing cancer cells in vitro and in vivo was developed. Here, an antibody–antigen complex consisting of the single-chain variable fragment (scFv) of chA21 and an N-terminal fragment (residues 1–192, named EP I) of the ErbB2 extracellular domain was crystallized using the sitting-drop vapour-diffusion method. An X-ray diffraction data set was collected to 2.45 Å resolution from a single flash-cooled crystal; the crystal belonged to space group P212121. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Liu, Y. Zhou, H. Zhu, J. Gao, Y. Niu, L. Liu, J. Teng, M. 2009-06-27 doi:10.1107/S1744309109020107 International Union of Crystallography An antibody–antigen complex consisting of a single-chain variable fragment of the potential therapeutic antibody chA21 and an N-terminal fragment (residues 1–192) of the human ErbB2 extracellular domain was expressed, purified and crystallized. X-ray diffraction data were collected to 2.45 Å resolution. en chA21 ErbB2 antibodies antigens ErbB2 is a transmembrane tyrosine kinase, the overexpression of which causes abnormality and disorder in cell signalling and leads to cell transformation. Previously, an anti-ErbB2 single-chain chimeric antibody chA21 that specifically inhibits the growth of ErbB2-overexpressing cancer cells in vitro and in vivo was developed. Here, an antibody–antigen complex consisting of the single-chain variable fragment (scFv) of chA21 and an N-terminal fragment (residues 1–192, named EP I) of the ErbB2 extracellular domain was crystallized using the sitting-drop vapour-diffusion method. An X-ray diffraction data set was collected to 2.45 Å resolution from a single flash-cooled crystal; the crystal belonged to space group P212121. text/html Crystallization and preliminary crystallographic studies of the single-chain variable fragment of antibody chA21 in complex with an N-terminal fragment of ErbB2 text 7 65 2009-06-27 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 692 694 http://scripts.iucr.org/cgi-bin/paper?fw5221 Two mutants of the toxic extracellular zinc endopeptidase AsaP1 (AsaP1_E294Q and AsaP1_E294A) of Aeromonas salmonicida subsp. achromogenes were expressed in Escherichia coli and crystallized by the vapour-diffusion method. Crystals were obtained using several precipitants and different protein concentrations. Protein crystals were found in a monoclinic (C2) as well as an orthorhombic (P212121) space group. The crystals belonging to the monoclinic space group C2 had unit-cell parameters a = 103.4, b = 70.9, c = 54.9 Å, β = 109.3° for AsaP1_E294A, and a = 98.5, b = 74.5, c = 54.7 Å, β = 112.4° for AsaP1_E294Q. The unit-cell parameters of the orthorhombic crystal obtained for AsaP1_E294A were a = 57.9, b = 60.2, c = 183.6 Å. The crystals of the two different mutants diffracted X-rays beyond 2.0 Å resolution. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Bogdanović, X. Singh, R.K. Hentschke, J. Gudmundsdóttir, B.K. Hinrichs, W. 2009-06-27 doi:10.1107/S1744309109020132 International Union of Crystallography Crystallization and preliminary X-ray diffraction studies of AsaP1_E294A and AsaP1_E294Q, two inactive mutants of the toxic zinc metallopeptidase AsaP1 from A. salmonicida subsp. achromogenes, are reported. en zinc metallopeptidases Aeromonas salmonicida subsp. achromogenes Two mutants of the toxic extracellular zinc endopeptidase AsaP1 (AsaP1_E294Q and AsaP1_E294A) of Aeromonas salmonicida subsp. achromogenes were expressed in Escherichia coli and crystallized by the vapour-diffusion method. Crystals were obtained using several precipitants and different protein concentrations. Protein crystals were found in a monoclinic (C2) as well as an orthorhombic (P212121) space group. The crystals belonging to the monoclinic space group C2 had unit-cell parameters a = 103.4, b = 70.9, c = 54.9 Å, β = 109.3° for AsaP1_E294A, and a = 98.5, b = 74.5, c = 54.7 Å, β = 112.4° for AsaP1_E294Q. The unit-cell parameters of the orthorhombic crystal obtained for AsaP1_E294A were a = 57.9, b = 60.2, c = 183.6 Å. The crystals of the two different mutants diffracted X-rays beyond 2.0 Å resolution. text/html Crystallization and preliminary X-ray diffraction studies of AsaP1_E294A and AsaP1_E294Q, two inactive mutants of the toxic zinc metallopeptidase AsaP1 from Aeromonas salmonicida subsp. achromogenes text 7 65 2009-06-27 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 695 697 http://scripts.iucr.org/cgi-bin/paper?ll5187 Glycerol dehydrogenase (GldA) encoded by the STM4108 gene (gldA) has been related to the synthesis of HilA, a major transcriptional regulator that is responsible for the expression of invasion genes in the human pathogen Salmonella enterica serovar Typhimurium. Single colourless crystals were obtained from a recombinant preparation of GldA overexpressed in Escherichia coli. They belonged to space group P2221, with unit-cell parameters a = 127.0, b = 160.1, c = 665.2 Å. The crystals contained a very large number of molecules in the asymmetric unit, probably 30–35. Diffraction data were collected to 3.5 Å resolution using synchrotron radiation at the European Synchrotron Radiation Facility. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Gonçalves, A.T. Marçal, D. Carrondo, M.A. Enguita, F.J. 2009-06-27 doi:10.1107/S1744309109020296 International Union of Crystallography The expression, purification and crystallization of a glycerol dehydrogenase from S. typhimurium is decribed. The crystals diffracted to 3.5 Å resolution. en glycerol dehydrogenase Salmonella enterica serovar Typhimurium family III alcohol dehydrogenases foodborne infections Glycerol dehydrogenase (GldA) encoded by the STM4108 gene (gldA) has been related to the synthesis of HilA, a major transcriptional regulator that is responsible for the expression of invasion genes in the human pathogen Salmonella enterica serovar Typhimurium. Single colourless crystals were obtained from a recombinant preparation of GldA overexpressed in Escherichia coli. They belonged to space group P2221, with unit-cell parameters a = 127.0, b = 160.1, c = 665.2 Å. The crystals contained a very large number of molecules in the asymmetric unit, probably 30–35. Diffraction data were collected to 3.5 Å resolution using synchrotron radiation at the European Synchrotron Radiation Facility. text/html Crystallization and preliminary X-ray characterization of a glycerol dehydrogenase from the human pathogen Salmonella enterica serovar Typhimurium text 7 65 2009-06-27 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 698 701 http://scripts.iucr.org/cgi-bin/paper?en5360 EpsH is a minor pseudopilin protein of the Vibrio cholerae type II secretion system. A truncated form of EpsH with a C-terminal noncleavable His tag was constructed and expressed in Escherichia coli, purified and crystallized by sitting-drop vapor diffusion. A complete data set was collected to 1.71 Å resolution. The crystals belonged to space group P212121, with unit-cell parameters a = 53.39, b = 71.11, c = 84.64 Å. There were two protein molecules in the asymmetric unit, which gave a Matthews coefficient VM of 2.1 Å3 Da−1, corresponding to 41.5% solvent content. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Raghunathan, K. Vago, F.S. Ball, T. Yakubova, N. Grindem, D. Wedemeyer, W.J. Arvidson, D.N. 2009-06-27 doi:10.1107/S1744309109020454 International Union of Crystallography Recombinant V. cholerae EpsH has been expressed, purified and crystallized. The crystals diffracted to 1.71 Å resolution. en EpsH GspH Vibrio cholerae type II secretion system pseudopilins pseudopilus EpsH is a minor pseudopilin protein of the Vibrio cholerae type II secretion system. A truncated form of EpsH with a C-terminal noncleavable His tag was constructed and expressed in Escherichia coli, purified and crystallized by sitting-drop vapor diffusion. A complete data set was collected to 1.71 Å resolution. The crystals belonged to space group P212121, with unit-cell parameters a = 53.39, b = 71.11, c = 84.64 Å. There were two protein molecules in the asymmetric unit, which gave a Matthews coefficient VM of 2.1 Å3 Da−1, corresponding to 41.5% solvent content. text/html Expression, purification, crystallization and preliminary X-ray studies of Vibrio choleraepseudopilin EpsH text 7 65 2009-06-27 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 702 704 http://scripts.iucr.org/cgi-bin/paper?en5365 7C8 is a mouse monoclonal antibody that is specific for the third hypervariable loop (V3 loop) of the human immunodeficiency virus type 2 (HIV-2) associated protein gp125. Fab fragments of 7C8 effectively neutralize HIV-2. 7C8 was expressed and purified from a hybridoma cell line in order to establish the molecular basis underlying the specificity of the 7C8 antibody for the V3 loop as well as the specific role of the elongated third complementarity-determining region of the heavy chain (CDRH3). The antibody was digested with papain and Fab fragments were purified using size-exclusion chromatography. Hanging-drop vapour-diffusion crystallization techniques were employed and the protein was crystallized in 50 mM ammonium sulfate, 100 mM Tris–HCl pH 8.5, 25%(w/v) PEG 8000 and 2.5%(w/v) PEG 400 at 275 K. The analysed crystals belonged to the rhombohedral space group P3221, with unit-cell parameters a = b = 100.1, c = 196.8 Å, and diffracted to 2.7 Å resolution. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Uchtenhagen, H. Sourial, S. Friemann, R. Ehnlund, M. Spetz, A.-L. Harris, R.A. Madhurantakam, C. Achour, A. 2009-06-27 doi:10.1107/S1744309109020685 International Union of Crystallography Neutralizing Fab fragments of the HIV-2-binding murine antibody 7C8 were generated after purification from hybridoma cell-culture supernatant. Crystallization conditions were determined and diffraction data were collected to 2.7 Å resolution. en 7C8 Fab fragment HIV antibodies 7C8 is a mouse monoclonal antibody that is specific for the third hypervariable loop (V3 loop) of the human immunodeficiency virus type 2 (HIV-2) associated protein gp125. Fab fragments of 7C8 effectively neutralize HIV-2. 7C8 was expressed and purified from a hybridoma cell line in order to establish the molecular basis underlying the specificity of the 7C8 antibody for the V3 loop as well as the specific role of the elongated third complementarity-determining region of the heavy chain (CDRH3). The antibody was digested with papain and Fab fragments were purified using size-exclusion chromatography. Hanging-drop vapour-diffusion crystallization techniques were employed and the protein was crystallized in 50 mM ammonium sulfate, 100 mM Tris–HCl pH 8.5, 25%(w/v) PEG 8000 and 2.5%(w/v) PEG 400 at 275 K. The analysed crystals belonged to the rhombohedral space group P3221, with unit-cell parameters a = b = 100.1, c = 196.8 Å, and diffracted to 2.7 Å resolution. text/html Production, purification, crystallization and preliminary X-ray diffraction analysis of the HIV-2-neutralizing V3 loop-specific Fab fragment 7C8 text 7 65 2009-06-27 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 705 708 http://scripts.iucr.org/cgi-bin/paper?hc5079 Glycinamide ribonucleotide transformylase (GART) catalyzes the transfer of a formyl group from formyl tetrahydrofolate (FTHF) to glycinamide ribonucleotide (GAR), which is an essential step in the de novo synthesis pathway of purines. In Bacillus subtilis, GART is encoded by the gene purN. In order to study the structure and function of B. subtilis GART, the purN gene was amplified, cloned into an expression vector and expressed in soluble form in Escherichia coli. The protein was purified to homogeneity and crystals suitable for X-ray data collection were obtained. These crystals diffracted to 2.5 Å resolution and belonged to space group P3121, with unit-cell parameters a = b = 95.5, c = 64.0 Å. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Liang, Y.-H. Liu, X.-Y. Wang, J. Li, L.-F. 2009-06-27 doi:10.1107/S1744309109020703 International Union of Crystallography In order to study the structure and function of B. subtilis glycinamide ribonucleotide transformylase, the purN gene was amplified, cloned into an expression vector and expressed in soluble form in Escherichia coli. The protein was purified to homogeneity and crystals suitable for X-ray data collection were obtained. en glycinamide ribonucleotide transformylase Bacillus subtilis purine synthesis Glycinamide ribonucleotide transformylase (GART) catalyzes the transfer of a formyl group from formyl tetrahydrofolate (FTHF) to glycinamide ribonucleotide (GAR), which is an essential step in the de novo synthesis pathway of purines. In Bacillus subtilis, GART is encoded by the gene purN. In order to study the structure and function of B. subtilis GART, the purN gene was amplified, cloned into an expression vector and expressed in soluble form in Escherichia coli. The protein was purified to homogeneity and crystals suitable for X-ray data collection were obtained. These crystals diffracted to 2.5 Å resolution and belonged to space group P3121, with unit-cell parameters a = b = 95.5, c = 64.0 Å. text/html Protein preparation, crystallization and preliminary crystallographic studies of Bacillus subtilisglycinamide ribonucleotide transformylase text 7 65 2009-06-27 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 709 711 http://scripts.iucr.org/cgi-bin/paper?bw5296 SspB is a 1500-residue adhesin expressed on the surface of the oral bacterium Streptococcus gordonii. Its interaction with other bacteria and host cells initiates the development of dental plaque. The full-length C-terminal domain of SspB was cloned, overexpressed in Escherichia coli and purified. However, the protein could not be crystallized. Limited proteolysis of the full-length C-domain identified a core fragment. The proteolysis product was cloned, expressed and purified. The protein was crystallized using the hanging-drop vapour-diffusion method. X-ray data were collected and processed to a maximum resolution of 2.1 Å with 96.4% completeness. The crystals belonged to space group P21, with one molecule in the asymmetric unit, a solvent content of 33.7% and a corresponding Matthews coefficient of 1.85 Å3 Da−1. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Forsgren, N. Lamont, R.J. Persson, K. 2009-06-27 doi:10.1107/S1744309109021046 International Union of Crystallography The C-terminal domain of the S. gordonii surface protein SspB has been expressed, purified and crystallized. Diffraction data have been collected to 2.1 Å resolution. en SspB C-terminal domain Streptococcus gordonii dental plaque SspB is a 1500-residue adhesin expressed on the surface of the oral bacterium Streptococcus gordonii. Its interaction with other bacteria and host cells initiates the development of dental plaque. The full-length C-terminal domain of SspB was cloned, overexpressed in Escherichia coli and purified. However, the protein could not be crystallized. Limited proteolysis of the full-length C-domain identified a core fragment. The proteolysis product was cloned, expressed and purified. The protein was crystallized using the hanging-drop vapour-diffusion method. X-ray data were collected and processed to a maximum resolution of 2.1 Å with 96.4% completeness. The crystals belonged to space group P21, with one molecule in the asymmetric unit, a solvent content of 33.7% and a corresponding Matthews coefficient of 1.85 Å3 Da−1. text/html A crystallizable form of the Streptococcus gordonii surface antigen SspB C-domain obtained by limited proteolysis text 7 65 2009-06-27 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 712 714 http://scripts.iucr.org/cgi-bin/paper?pu5264 Adeno-associated virus (AAV) serotype 9, which is under development for gene-delivery applications, shows significantly enhanced capsid-associated transduction efficiency in muscle compared with other AAV serotypes. With the aim of characterizing the structural determinants of this property, the purification, crystallization and preliminary X-ray crystallographic analyses of the AAV9 viral capsid are reported. The crystals diffracted X-rays to 2.8 Å resolution using synchrotron radiation and belonged to the trigonal space group P32, with unit-cell parameters a = b = 251.0, c = 640.0 Å. There are three complete viral capsids in the crystal unit cell. The orientation and position of the asymmetric unit capsid have been determined by molecular-replacement methods and structure determination is in progress. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Mitchell, M. Nam, H.-J. Carter, A. McCall, A. Rence, C. Bennett, A. Gurda, B. McKenna, R. Porter, M. Sakai, Y. Byrne, B.J. Muzyczka, N. Aslanidi, G. Zolotukhin, S. Agbandje-McKenna, M. 2009-06-27 doi:10.1107/S1744309109021460 International Union of Crystallography The purification, crystallization and preliminary X-ray crystallographic analyses of the AAV9 viral capsid are reported. en adeno-associate virus serotype 9 viruses Adeno-associated virus (AAV) serotype 9, which is under development for gene-delivery applications, shows significantly enhanced capsid-associated transduction efficiency in muscle compared with other AAV serotypes. With the aim of characterizing the structural determinants of this property, the purification, crystallization and preliminary X-ray crystallographic analyses of the AAV9 viral capsid are reported. The crystals diffracted X-rays to 2.8 Å resolution using synchrotron radiation and belonged to the trigonal space group P32, with unit-cell parameters a = b = 251.0, c = 640.0 Å. There are three complete viral capsids in the crystal unit cell. The orientation and position of the asymmetric unit capsid have been determined by molecular-replacement methods and structure determination is in progress. text/html Production, purification and preliminary X-ray crystallographic studies of adeno-associated virus serotype 9 text 7 65 2009-06-27 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 715 718 http://scripts.iucr.org/cgi-bin/paper?rp5034 YycGF is a crucial signal transduction system for the regulation of cell-wall metabolism in low-G+C Gram-positive bacteria, which include many important human pathogens. The response regulator YycF receives signals from its cognate histidine kinase YycG through a phosphotransfer reaction and elicits responses through regulation of gene expression. The N-terminal regulatory domain of YycF from Bacillus subtilis was overproduced and purified. The protein was crystallized and X-ray data were collected to 1.95 Å resolution with a completeness of 97.7% and an overall Rmerge of 7.7%. The crystals belonged to space group P3121, with unit-cell parameters a = b = 59.50, c = 79.06 Å. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Zhao, H. Heroux, A. Sequeira, R.D. Tang, L. 2009-06-27 doi:10.1107/S1744309109022696 International Union of Crystallography The regulatory domain of response regulator YycF from an essential two-component signal transduction system has been crystallized and X-ray data have been collected at 1.95 Å resolution. en response regulators two-component systems signal transduction Gram-positive bacteria YycF YycG transcription factor cell wall YycGF is a crucial signal transduction system for the regulation of cell-wall metabolism in low-G+C Gram-positive bacteria, which include many important human pathogens. The response regulator YycF receives signals from its cognate histidine kinase YycG through a phosphotransfer reaction and elicits responses through regulation of gene expression. The N-terminal regulatory domain of YycF from Bacillus subtilis was overproduced and purified. The protein was crystallized and X-ray data were collected to 1.95 Å resolution with a completeness of 97.7% and an overall Rmerge of 7.7%. The crystals belonged to space group P3121, with unit-cell parameters a = b = 59.50, c = 79.06 Å. text/html Preliminary crystallographic studies of the regulatory domain of response regulator YycF from an essential two-component signal transduction system text 7 65 2009-06-27 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 719 722 http://scripts.iucr.org/cgi-bin/paper?en5367 Human SOUL (hSOUL) is a 23 kDa haem-binding protein that was first identified as the PP23 protein isolated from human full-term placentas. Here, the overexpression, purification and crystallization of hSOUL are reported. The crystals belonged to space group P6422, with unit-cell parameters a = b = 145, c = 60 Å and one protein molecule in the asymmetric unit. X-ray diffraction data were collected to 3.5 Å resolution at the ESRF. A preliminary model of the three-dimensional structure of hSOUL was obtained by molecular replacement using the structures of murine p22HBP (PDB codes 2gov and 2hva), obtained by solution NMR, as search models. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Freire, F. Romão, M.J. Macedo, A.L. Aveiro, S.S. Goodfellow, B.J. Carvalho, A.L. 2009-06-27 doi:10.1107/S174430910902291X International Union of Crystallography This manuscript describes the overexpression, purification and crystallization of human SOUL protein (hSOUL). hSOUL is a 23 kDa haem-binding protein that was first identified as the PP23 protein isolated from human full-term placenta. en haem-binding proteins SOUL Human SOUL (hSOUL) is a 23 kDa haem-binding protein that was first identified as the PP23 protein isolated from human full-term placentas. Here, the overexpression, purification and crystallization of hSOUL are reported. The crystals belonged to space group P6422, with unit-cell parameters a = b = 145, c = 60 Å and one protein molecule in the asymmetric unit. X-ray diffraction data were collected to 3.5 Å resolution at the ESRF. A preliminary model of the three-dimensional structure of hSOUL was obtained by molecular replacement using the structures of murine p22HBP (PDB codes 2gov and 2hva), obtained by solution NMR, as search models. text/html Preliminary structural characterization of human SOUL, a haem-binding protein text 7 65 2009-06-27 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 723 726 http://scripts.iucr.org/cgi-bin/paper?nj5037 The response regulator DesR, which activates the transcription of the des gene by binding to a regulatory region, is essential for controlling the fluidity of membrane phospholipids. DesR from Streptococcus pneumoniae was overexpressed in Escherichia coli. The protein was purified and crystallized for structural analysis. Diffraction data were collected to 1.7 Å resolution using synchrotron radiation and the crystals belonged to the orthorhombic space group P212121, with unit-cell parameters a = 46.91, b = 71.38, c = 117.73 Å. Assuming the presence of a dimer in the asymmetric unit, this corresponds to a VM of 2.21 Å3 Da−1. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Park, A.K. Bong, S.M. Moon, J.H. Chi, Y.M. 2009-06-27 doi:10.1107/S1744309109023082 International Union of Crystallography The response regulator DesR from S. pneumoniae was cloned, expressed, purified and crystallized. A complete data set was collected to 1.7 Å resolution. en two-component systems fatty-acid desaturation response regulators The response regulator DesR, which activates the transcription of the des gene by binding to a regulatory region, is essential for controlling the fluidity of membrane phospholipids. DesR from Streptococcus pneumoniae was overexpressed in Escherichia coli. The protein was purified and crystallized for structural analysis. Diffraction data were collected to 1.7 Å resolution using synchrotron radiation and the crystals belonged to the orthorhombic space group P212121, with unit-cell parameters a = 46.91, b = 71.38, c = 117.73 Å. Assuming the presence of a dimer in the asymmetric unit, this corresponds to a VM of 2.21 Å3 Da−1. text/html Crystallization and preliminary X-ray crystallographic studies of DesR, a thermosensing response regulator in a two-component signalling system from Streptococcus pneumoniae text 7 65 2009-06-27 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 727 729 http://scripts.iucr.org/cgi-bin/paper?gj5064 The orange-coloured protein (ORP) from Desulfovibrio gigas is a 12 kDa protein that contains a novel mixed-metal sulfide cluster of the type [S2MoS2CuS2MoS2]. Diffracting crystals of the apo form of ORP have been obtained. Data have been collected for the apo form of ORP to 2.25 Å resolution in-house and to beyond 2.0 Å resolution at ESRF, Grenoble. The crystals belonged to a trigonal space group, with unit-cell parameters a = 43, b = 43, c = 106 Å. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Najmudin, S. Bonifácio, C. Duarte, A.G. Pualeta, S.R. Moura, I. Moura, J.J.G. Romão, M.J. 2009-06-30 doi:10.1107/S1744309109023392 International Union of Crystallography Diifraction data have been collected for the apo form of the orange protein from D. gigas to 2.25 Å resolution in-house and to beyond 2.0 Å resolution at ESRF, Grenoble. The crystals belonged to a trigonal space group, with unit-cell parameters a = 43, b = 43, c = 106 Å. en mixed-metal sulfide cluster Desulfovibrio gigas molybdenum copper The orange-coloured protein (ORP) from Desulfovibrio gigas is a 12 kDa protein that contains a novel mixed-metal sulfide cluster of the type [S2MoS2CuS2MoS2]. Diffracting crystals of the apo form of ORP have been obtained. Data have been collected for the apo form of ORP to 2.25 Å resolution in-house and to beyond 2.0 Å resolution at ESRF, Grenoble. The crystals belonged to a trigonal space group, with unit-cell parameters a = 43, b = 43, c = 106 Å. text/html Crystallization and crystallographic analysis of the apo form of the orange protein (ORP) from Desulfovibrio gigas text 7 65 2009-06-30 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 730 732 http://scripts.iucr.org/cgi-bin/paper?en5369 The enzyme phosphoglucosamine mutase catalyzes the conversion of glucosamine 6-phosphate to glucosamine 1-phosphate, an early step in the formation of the nucleotide sugar UDP-N-acetylglucosamine, which is involved in peptidoglycan biosynthesis. These enzymes are part of the large α-d-phosphohexomutase enzyme superfamily, but no proteins from the phosphoglucosamine mutase subgroup have been structurally characterized to date. Here, the crystallization of phosphoglucosamine mutase from Bacillus anthracis in space group P3221 by hanging-drop vapor diffusion is reported. The crystals diffracted to 2.7 Å resolution under cryocooling conditions. Structure determination by molecular replacement was successful and refinement is under way. The crystal structure of B. anthracis phosphoglucosamine mutase should shed light on the substrate-specificity of these enzymes and will also serve as a template for inhibitor design. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Mehra-Chaudhary, R. Neace, C.E. Beamer, L.J. 2009-06-30 doi:10.1107/S1744309109023409 International Union of Crystallography The enzyme phosphoglucosamine mutase from B. anthracis participates in the peptidoglycan-biosynthetic pathway. The expression, purification and crystallization of this enzyme are described; diffraction data have been collected to 2.7 Å resolution. en phosphoglucosamine mutase Bacillus anthracis peptidoglycan biosynthesis The enzyme phosphoglucosamine mutase catalyzes the conversion of glucosamine 6-phosphate to glucosamine 1-phosphate, an early step in the formation of the nucleotide sugar UDP-N-acetylglucosamine, which is involved in peptidoglycan biosynthesis. These enzymes are part of the large α-d-phosphohexomutase enzyme superfamily, but no proteins from the phosphoglucosamine mutase subgroup have been structurally characterized to date. Here, the crystallization of phosphoglucosamine mutase from Bacillus anthracis in space group P3221 by hanging-drop vapor diffusion is reported. The crystals diffracted to 2.7 Å resolution under cryocooling conditions. Structure determination by molecular replacement was successful and refinement is under way. The crystal structure of B. anthracis phosphoglucosamine mutase should shed light on the substrate-specificity of these enzymes and will also serve as a template for inhibitor design. text/html Crystallization and initial crystallographic analysis of phosphoglucosamine mutase from Bacillus anthracis text 7 65 2009-06-30 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 733 735 http://scripts.iucr.org/cgi-bin/paper?ll5190 A Kunitz-type proteinase inhibitor has been purified from tamarind (Tamarindus indica) seeds. SDS–PAGE analysis of a purified sample showed a homogeneous band corresponding to a molecular weight of 21 kDa. The protein was identified as a Kunitz-type proteinase inhibitor based on N-terminal amino-acid sequence analysis. It was crystallized by the vapour-diffusion method using PEG 6000. The crystals belonged to the orthorhombic space group C2221, with unit-cell parameters a = 37.2, b = 77.1, c = 129.1 Å. Diffraction data were collected to a resolution of 2.7 Å. Preliminary crystallographic analysis indicated the presence of one proteinase inhibitor molecule in the asymmetric unit, with a solvent content of 44%. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Patil, D.N. Preeti Chaudhry, A. Sharma, A.K. Tomar, S. Kumar, P. 2009-06-30 doi:10.1107/S1744309109023495 International Union of Crystallography A 21 kDa Kunitz-type proteinase inhibitor was purified from tamarind (T. indica) seeds, crystallized and characterized by X-ray diffraction. en Kunitz-type proteinase inhibitors Tamarindus indica A Kunitz-type proteinase inhibitor has been purified from tamarind (Tamarindus indica) seeds. SDS–PAGE analysis of a purified sample showed a homogeneous band corresponding to a molecular weight of 21 kDa. The protein was identified as a Kunitz-type proteinase inhibitor based on N-terminal amino-acid sequence analysis. It was crystallized by the vapour-diffusion method using PEG 6000. The crystals belonged to the orthorhombic space group C2221, with unit-cell parameters a = 37.2, b = 77.1, c = 129.1 Å. Diffraction data were collected to a resolution of 2.7 Å. Preliminary crystallographic analysis indicated the presence of one proteinase inhibitor molecule in the asymmetric unit, with a solvent content of 44%. text/html Purification, crystallization and preliminary crystallographic studies of a Kunitz-type proteinase inhibitor from tamarind (Tamarindus indica) seeds text 7 65 2009-06-30 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 736 738 http://scripts.iucr.org/cgi-bin/paper?ll5193 ADAMTS13 is a reprolysin-type metalloproteinase belonging to the ADAMTS (a disintegrin and metalloproteinase with thrombospondin type 1 motif) family. It specifically cleaves plasma von Willebrand factor (VWF) and regulates platelet adhesion and aggregation. ADAMTS13 is a multi-domain enzyme. In addition to the N-terminal metalloproteinase domain, the ancillary domains, including a disintegrin-like domain, a thrombospondin-1 type 1 repeat, a Cys-rich domain and a spacer domain, are required for VWF recognition and cleavage. In the present study, a fragment of the ADAMTS13 ancillary domains (ADAMTS13-DTCS; residues 287–685) was expressed using CHO Lec cells, purified and crystallized. Diffraction data sets were collected using the SPring-8 beamline. Two ADAMTS13-DTCS crystals with distinct unit-cell parameters generated data sets to 2.6 and 2.8 Å resolution, respectively. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Akiyama, M. Takeda, S. Kokame, K. Takagi, J. Miyata, T. 2009-06-30 doi:10.1107/S1744309109023410 International Union of Crystallography A fragment of the ADAMTS13 ancillary domains (ADAMTS13-DTCS) has been expressed, purified and crystallized and the crystals have been characterized by X-ray diffraction. en von Willebrand factor-cleaving proteinase ADAMTS13 metalloproteinases ancillary domains ADAMTS13 is a reprolysin-type metalloproteinase belonging to the ADAMTS (a disintegrin and metalloproteinase with thrombospondin type 1 motif) family. It specifically cleaves plasma von Willebrand factor (VWF) and regulates platelet adhesion and aggregation. ADAMTS13 is a multi-domain enzyme. In addition to the N-terminal metalloproteinase domain, the ancillary domains, including a disintegrin-like domain, a thrombospondin-1 type 1 repeat, a Cys-rich domain and a spacer domain, are required for VWF recognition and cleavage. In the present study, a fragment of the ADAMTS13 ancillary domains (ADAMTS13-DTCS; residues 287–685) was expressed using CHO Lec cells, purified and crystallized. Diffraction data sets were collected using the SPring-8 beamline. Two ADAMTS13-DTCS crystals with distinct unit-cell parameters generated data sets to 2.6 and 2.8 Å resolution, respectively. text/html Production, crystallization and preliminary crystallographic analysis of an exosite-containing fragment of human von Willebrand factor-cleaving proteinase ADAMTS13 text 7 65 2009-06-30 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 739 742 http://scripts.iucr.org/cgi-bin/paper?bw5299 Periplasmic membrane-fusion proteins (MFPs) are an essential component of multidrug and metal-efflux pumps in Gram-negative bacteria. However, the functional structure of MFPs remains unclear. CusCFBA, the CuI and AgI efflux system in Escherichia coli, consists of the MFP CusB, the OMF CusC and the RND-type transporter CusA. The MFP CusB bridges the inner membrane RND-type efflux transporter CusA and the outer membrane factor CusC and exhibits substrate-linked conformational changes which distinguish it from other MFP-family members. CusB from E. coli was overexpressed and the recombinant protein was purified using Ni–NTA affinity, Q anion-exchange and gel-filtration chromatography. The purified CusB protein was crystallized using the vapour-diffusion method. A diffraction data set was collected to a resolution of 3.1 Å at 100 K. The crystal belonged to space group C222. Copyright (c) 2009 International Union of Crystallography urn:issn:1744-3091 Xu, Y. Yun, B.-Y. Sim, S.-H. Lee, K. Ha, N.-C. 2009-06-30 doi:10.1107/S1744309109019873 International Union of Crystallography This article describes how CusB from E. coli was overexpressed and the recombinant protein purified using Ni–NTA affinity, Q anion-exchange and gel-filtration chromatography, and how the purified CusB protein was crystallized using the vapour-diffusion method. en membrane-fusion proteins metal-efflux pumps Gram-negative bacteria RND-type transporters Periplasmic membrane-fusion proteins (MFPs) are an essential component of multidrug and metal-efflux pumps in Gram-negative bacteria. However, the functional structure of MFPs remains unclear. CusCFBA, the CuI and AgI efflux system in Escherichia coli, consists of the MFP CusB, the OMF CusC and the RND-type transporter CusA. The MFP CusB bridges the inner membrane RND-type efflux transporter CusA and the outer membrane factor CusC and exhibits substrate-linked conformational changes which distinguish it from other MFP-family members. CusB from E. coli was overexpressed and the recombinant protein was purified using Ni–NTA affinity, Q anion-exchange and gel-filtration chromatography. The purified CusB protein was crystallized using the vapour-diffusion method. A diffraction data set was collected to a resolution of 3.1 Å at 100 K. The crystal belonged to space group C222. text/html Crystallization and preliminary X-ray crystallographic analysis of Escherichia coli CusB text 7 65 2009-06-30 Copyright (c) 2009 International Union of Crystallography Acta Crystallographica Section F: Structural Biology and Crystallization Communications crystallization communications 743 745