4ALE

Structure changes of Polysaccharide monooxygenase CBM33A from Enterococcus faecalis by X-ray induced photoreduction.

4ALE の概要

• エントリーDOI: 10.2210/pdb4ale/pdb
• 関連するPDBエントリー: 4A02 4ALC 4ALQ 4ALR 4ALS 4ALT
• 分子名称: CHITIN BINDING PROTEIN, COPPER (II) ION, DI(HYDROXYETHYL)ETHER, ... (4 entities in total)
• 機能のキーワード: chitin binding protein, cbm33, chitin degradation, microspectrophotometry
• 由来する生物種: ENTEROCOCCUS FAECALIS
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 18721.21

構造登録者

Gudmundsson, M.,Wu, M.,Ishida, T.,Momeni, M.H.,Vaaje-Kolstad, G.,Eijsink, V.,Sandgren, M. (登録日: 2012-03-02, 公開日: 2013-02-27, 最終更新日: 2023-12-20)

主引用文献

Gudmundsson, M.,Kim, S.,Wu, M.,Ishida, T.,Haddad Momeni, M.,Vaaje-Kolstad, G.,Lundberg, D.,Royant, A.,Stahlberg, J.,Eijsink, V.G.,Beckham, G.T.,Sandgren, M.
Structural and Electronic Snapshots During the Transition from a Cu(II) to Cu(I) Metal Center of a Lytic Polysaccharide Monooxygenase by X-Ray Photo-Reduction.
J.Biol.Chem., 289:18782-, 2014

PubMed Abstract: Lytic polysaccharide monooxygenases (LPMOs) are a recently discovered class of enzymes that employ a copper-mediated, oxidative mechanism to cleave glycosidic bonds. The LPMO catalytic mechanism likely requires that molecular oxygen first binds to Cu(I), but the oxidation state in many reported LPMO structures is ambiguous, and the changes in the LPMO active site required to accommodate both oxidation states of copper have not been fully elucidated. Here, a diffraction data collection strategy minimizing the deposited x-ray dose was used to solve the crystal structure of a chitin-specific LPMO from Enterococcus faecalis (EfaCBM33A) in the Cu(II)-bound form. Subsequently, the crystalline protein was photoreduced in the x-ray beam, which revealed structural changes associated with the conversion from the initial Cu(II)-oxidized form with two coordinated water molecules, which adopts a trigonal bipyramidal geometry, to a reduced Cu(I) form in a T-shaped geometry with no coordinated water molecules. A comprehensive survey of Cu(II) and Cu(I) structures in the Cambridge Structural Database unambiguously shows that the geometries observed in the least and most reduced structures reflect binding of Cu(II) and Cu(I), respectively. Quantum mechanical calculations of the oxidized and reduced active sites reveal little change in the electronic structure of the active site measured by the active site partial charges. Together with a previous theoretical investigation of a fungal LPMO, this suggests significant functional plasticity in LPMO active sites. Overall, this study provides molecular snapshots along the reduction process to activate the LPMO catalytic machinery and provides a general method for solving LPMO structures in both copper oxidation states.

PubMed: 24828494
DOI: 10.1074/JBC.M114.563494

実験手法

X-RAY DIFFRACTION (1.48 Å)