3WWD

The complex of pOPH_S172C with DMSO

3WWD の概要

• エントリーDOI: 10.2210/pdb3wwd/pdb
• 分子名称: Oxidized polyvinyl alcohol hydrolase, CITRIC ACID, DIMETHYL SULFOXIDE, ... (4 entities in total)
• 機能のキーワード: tryptophan, disulfide bridge, p-nitrophenyl esters, hydrolase
• 由来する生物種: Pseudomonas sp. VM15C
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 39599.05

構造登録者

Yang, Y.,Ko, T.P.,Li, J.H.,Liu, L.,Huang, C.H.,Chen, J.,Guo, R.T.,Du, G.C. (登録日: 2014-06-17, 公開日: 2015-04-29, 最終更新日: 2023-12-06)

主引用文献

Yang, Y.,Ko, T.P.,Liu, L.,Li, J.,Huang, C.H.,Chen, J.,Guo, R.T.,Du, G.
Roles of tryptophan residue and disulfide bond in the variable lid region of oxidized polyvinyl alcohol hydrolase
Biochem.Biophys.Res.Commun., 452:509-514, 2014

PubMed Abstract: Oxidized polyvinyl alcohol hydrolase (OPH) catalyzes the cleavage of C-C bond in β-diketone. It belongs to the α/β-hydrolase family and contains a unique lid region that covers the active site. The lid is the most variable region when pOPH from Pseudomonas sp. VM15C and sOPH from Sphingopyxis sp. 113P3 are compared. The wild-type enzymes and the pOPH mutants W255A, W255Y and W255F were analyzed for lipase activity by using p-nitrophenyl (pNP) esters as the substrates. The wild-type enzymes showed increased Km and decreased kcat/Km with the acyl chain length, and the mutants showed reduced kcat/Km for pNP acetate, indicating the importance of Trp255 in sequestering the active site from solvent. The significantly lower activity for pNP butyrate can be a result of product inhibition, as suggested by the complex crystal structures, in which butyric acid, DMSO or PEG occupied the same substrate-binding cleft. The mutant activity was retained with pNP caprylate and pNP laurate as the substrates, reflecting the amphipathic nature of the cleft. Moreover, the disulfide bond formation of Cys257/267 is important for the activity of pOPH, but it is not essential for sOPH, which has a shorter lid structure.

PubMed: 25173935
DOI: 10.1016/j.bbrc.2014.08.106

実験手法

X-RAY DIFFRACTION (1.65 Å)