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![[Figure 3]](di5060fig3mag.jpg)
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Figure 3
(a) AKG binding site comparison of in-line form (right) and off-line form (left). UbPH (silver), GriE (wheat, PDB code 5nci), fumitremorgin synthetase Ftmox1 (wheat, PDB code 4y5s; Yan et al., 2015  ), cisMIP4H (pink, PDB code 4p7w), arginine hydroxylase VioC (cyan, PDB code 6alm; Yin & Zabriskie, 2004) and taurine/AKG dioxygenase TauD (cyan, PDB code 1gqw). All ligands are shown in yellow sticks. The characteristic Gln in Ftmox1, anditomin biosynthesis enzyme (AndA) and ergot alkaloid (cycloclavine) synthase (ajEasH) are conserved in sequence, and there is an angle of 90° with the characteristic Asn/Gln in hydroxylases such as UbPH in spatial structure. This indicates that the conservativeness of the Asn/Gln in three-dimensional space may be related to the function of the protein. (b) Sequence alignment of βI sheets of AKG-dependent hydroxylases depends on structural alignment. The key residues are displayed in orange and blue. Clavaminic acid synthase Streptomyces clavuligerus (scCAS) (PDB code 1drt; Zhang et al., 2000), VioC (PDB code 6alm; Yin & Zabriskie, 2004), noramycin isomerase Streptomyces nogalater (SnoN) (PDB code 5equ; Siitonen et al., 2016), TauD (PDB code 1gqw; Elkins et al., 2002) and human hypoxia-inducible factor inhibitor asparaginyl hydroxylase (humanFI, FIH) (PDB code 1mzf; Dann et al., 2002) aligned with cisMIP4H. GriE (PDB code 5nci; Lukat et al., 2017), anthocyanin synthase Arabidopsis thaliana (atANS) (PDB code 1gp4; Wilmouth et al., 2002), vCPH (PDB code 5c5t; Longbotham et al., 2015), Ftmox1 (PDB code 4y5t; Yan et al., 2015), AndA (PDB code 5zm2; Nakashima et al., 2018) and ajEasH (PDB code 5m0t; Jakubczyk et al., 2016) aligned with UbPH. (c) Catalysis results of UbPH-wildtype and UbPH-N101I for 20 min reaction. An enzyme activity curve of UbPH-apo and UbPH-N101I (left). Values represent the mean ± s.d. of triplicates from a representative experiment (n = 3 experiment replicates), and the control value was subtracted. ND refers to a 4hyp concentration of less than 2. ***P<0.001 versus UbPH-N101I (right). One-way analysis of variance was used for data analysis using GraphPad Prism 6. |
![[Figure 3]](di5060fig3.jpg)
 | STRUCTURAL BIOLOGY |
ISSN: 2059-7983
