3C6X

HNL from Hevea brasiliensis to atomic resolution

3C6X の概要

• エントリーDOI: 10.2210/pdb3c6x/pdb
• 関連するPDBエントリー: 1QJ4 3C6Y 3C6Z 3C70 7YAS
• 分子名称: Hydroxynitrilase, SULFATE ION, BETA-MERCAPTOETHANOL, ... (5 entities in total)
• 機能のキーワード: atomic resolution, hydroxynitril lyase, catalysis, protonation state, ab initio calculations, substrate binding, lyase
• 由来する生物種: Hevea brasiliensis (Para rubber tree)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 29927.17

構造登録者

Schmidt, A. (登録日: 2008-02-06, 公開日: 2008-06-03, 最終更新日: 2023-11-01)

主引用文献

Schmidt, A.,Gruber, K.,Kratky, C.,Lamzin, V.S.
Atomic resolution crystal structures and quantum chemistry meet to reveal subtleties of hydroxynitrile lyase catalysis
J.Biol.Chem., 283:21827-21836, 2008

PubMed Abstract: Hydroxynitrile lyases are versatile enzymes that enantiospecifically cope with cyanohydrins, important intermediates in the production of various agrochemicals or pharmaceuticals. We determined four atomic resolution crystal structures of hydroxynitrile lyase from Hevea brasiliensis: one native and three complexes with acetone, isopropyl alcohol, and thiocyanate. We observed distinct distance changes among the active site residues related to proton shifts upon substrate binding. The combined use of crystallography and ab initio quantum chemical calculations allowed the determination of the protonation states in the enzyme active site. We show that His(235) of the catalytic triad must be protonated in order for catalysis to proceed, and we could reproduce the cyanohydrin synthesis in ab initio calculations. We also found evidence for the considerable pK(a) shifts that had been hypothesized earlier. We envision that this knowledge can be used to enhance the catalytic properties and the stability of the enzyme for industrial production of enantiomerically pure cyanohydrins.

PubMed: 18524775
DOI: 10.1074/jbc.M801056200

実験手法

X-RAY DIFFRACTION (1.05 Å)