2UYB

S161A mutant of Bacillus subtilis Oxalate Decarboxylase OxdC

2UYB の概要

• エントリーDOI: 10.2210/pdb2uyb/pdb
• 関連するPDBエントリー: 1J58 1L3J 1UW8 2UY9 2UYA
• 分子名称: OXALATE DECARBOXYLASE OXDC, FORMIC ACID, MANGANESE (II) ION, ... (5 entities in total)
• 機能のキーワード: lyase, cupin, formate, oxalate, manganese, s161a mutant, metal-binding, decarboxylase, metal binding protein
• 由来する生物種: BACILLUS SUBTILIS
• 細胞内の位置: Cytoplasm : O34714
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 43882.95

構造登録者

Just, V.J.,Burrell, M.R.,Bowater, L.,McRobbie, I.,Stevenson, C.E.M.,Lawson, D.M.,Bornemann, S. (登録日: 2007-04-03, 公開日: 2007-08-21, 最終更新日: 2023-12-13)

主引用文献

Just, V.J.,Burrell, M.R.,Bowater, L.,Mcrobbie, I.,Stevenson, C.E.M.,Lawson, D.M.,Bornemann, S.
The Identity of the Active Site of Oxalate Decarboxylase and the Importance of the Stability of Active-Site Lid Conformations.
Biochem.J., 407:397-, 2007

PubMed Abstract: Oxalate decarboxylase (EC 4.1.1.2) catalyses the conversion of oxalate into carbon dioxide and formate. It requires manganese and, uniquely, dioxygen for catalysis. It forms a homohexamer and each subunit contains two similar, but distinct, manganese sites termed sites 1 and 2. There is kinetic evidence that only site 1 is catalytically active and that site 2 is purely structural. However, the kinetics of enzymes with mutations in site 2 are often ambiguous and all mutant kinetics have been interpreted without structural information. Nine new site-directed mutants have been generated and four mutant crystal structures have now been solved. Most mutants targeted (i) the flexibility (T165P), (ii) favoured conformation (S161A, S164A, D297A or H299A) or (iii) presence (Delta162-163 or Delta162-164) of a lid associated with site 1. The kinetics of these mutants were consistent with only site 1 being catalytically active. This was particularly striking with D297A and H299A because they disrupted hydrogen bonds between the lid and a neighbouring subunit only when in the open conformation and were distant from site 2. These observations also provided the first evidence that the flexibility and stability of lid conformations are important in catalysis. The deletion of the lid to mimic the plant oxalate oxidase led to a loss of decarboxylase activity, but only a slight elevation in the oxalate oxidase side reaction, implying other changes are required to afford a reaction specificity switch. The four mutant crystal structures (R92A, E162A, Delta162-163 and S161A) strongly support the hypothesis that site 2 is purely structural.

PubMed: 17680775
DOI: 10.1042/BJ20070708

実験手法

X-RAY DIFFRACTION (2.1 Å)