3PL8

Pyranose 2-oxidase H167A complex with 3-deoxy-3-fluoro-beta-D-glucose

3PL8 の概要

• エントリーDOI: 10.2210/pdb3pl8/pdb
• 関連するPDBエントリー: 1TT0 2IGO
• 分子名称: Pyranose 2-oxidase, FLAVIN-ADENINE DINUCLEOTIDE, 2-(N-MORPHOLINO)-ETHANESULFONIC ACID, ... (5 entities in total)
• 機能のキーワード: substrate complex, h167a mutant, homotetramer, gmc oxidoreductase, phbh fold, rossmann domain, oxidoreductase, oxidase, fad cofactor, pyranose oxidation
• 由来する生物種: Trametes ochracea (White-rot fungus)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 70509.64

構造登録者

Tan, T.C.,Divne, C. (登録日: 2010-11-14, 公開日: 2011-04-27, 最終更新日: 2023-09-06)

主引用文献

Tan, T.C.,Haltrich, D.,Divne, C.
Regioselective control of Beta-d-glucose oxidation by pyranose 2-oxidase is intimately coupled to conformational degeneracy
J.Mol.Biol., 409:588-600, 2011

PubMed Abstract: Trametes multicolor pyranose 2-oxidase (P2O) is a flavoprotein oxidase that oxidizes d-glucose at C2 to 2-keto-d-glucose by a highly regioselective mechanism. In this work, fluorinated sugar substrates were used as mechanistic probes to investigate the basis of regioselectivity in P2O. Although frequently used to study the mechanisms of glycoside hydrolases, our work provides the first example of applying these probes to sugar oxidoreductases. Our previous structure of the P2O mutant H167A in complex with the slow substrate 2-deoxy-2-fluoro-d-glucose showed a substrate-binding mode compatible with oxidation at C3. To accommodate the sugar, a gating segment, (454)FSY(456), in the substrate recognition loop partly unfolded to create a spacious and more polar active site that is distinct from the closed state of P2O. The crystal structure presented here shows that the preferred C2 oxidation where an ordered complex of P2O H167A with 3-deoxy-3-fluoro-d-glucose at 1.35 Å resolution was successfully trapped. In this semi-open C2-oxidation complex, the substrate recognition loop tightens to form an optimized substrate complex stabilized by interactions between Asp452 and glucose O4, as well as Tyr456 and the glucose O6 group, interactions that are not possible when glucose is positioned for oxidation at C3. The different conformations of the (454)FSY(456) gating segment in the semi-open and closed states induce backbone and side-chain movements of Thr169 and Asp452 that add further differential stabilization to the individual states. We expect the semi-open state (C2-oxidation state) and closed state to be good approximations of the active-site structure during the reductive half-reaction (sugar oxidation) and oxidative half-reaction (O(2) reduction).

PubMed: 21515286
DOI: 10.1016/j.jmb.2011.04.019

実験手法

X-RAY DIFFRACTION (1.35 Å)