4YAS

HYDROXYNITRILE LYASE COMPLEXED WITH CHLORALHYDRATE

4YAS の概要

• エントリーDOI: 10.2210/pdb4yas/pdb
• 分子名称: PROTEIN (HYDROXYNITRILE LYASE), SULFATE ION, TRI-CHLORO-ACETALDEHYDE, ... (4 entities in total)
• 機能のキーワード: oxynitrilase, cyanogenesis, cyanohydrin formation, lyase
• 由来する生物種: Hevea brasiliensis
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 29698.17

構造登録者

Zuegg, J.,Wagner, U.G.,Gugganig, M.,Kratky, C. (登録日: 1999-03-15, 公開日: 1999-10-13, 最終更新日: 2024-11-20)

主引用文献

Zuegg, J.,Gruber, K.,Gugganig, M.,Wagner, U.G.,Kratky, C.
Three-dimensional structures of enzyme-substrate complexes of the hydroxynitrile lyase from Hevea brasiliensis.
Protein Sci., 8:1990-2000, 1999

PubMed Abstract: The 3D structures of complexes between the hydroxynitrile lyase from Hevea brasiliensis (Hb-HNL) and several substrate and/or inhibitor molecules, including trichloracetaldehyde, hexafluoracetone, acetone, and rhodanide, were determined by X-ray crystallography. The complex with trichloracetaldehyde showed a covalent linkage between the protein and the inhibitor, which had apparently resulted from nucleophilic attack of the catalytic Ser80-Ogamma. All other complexes showed the substrate or inhibitor molecule merely hydrogen bonded to the protein. In addition, the native crystal structure of Hb-HNL was redetermined at cryo-temperature and at room temperature, eliminating previous uncertainties concerning residual electron density within the active site, and leading to the observation of two conserved water molecules. One of them was found to be conserved in all complex structures and appears to have mainly structural significance. The other water molecule is conserved in all structures except for the complex with rhodanide; it is hydrogen bonded to the imidazole of the catalytic His235 and appears to affect the Hb-HNL catalyzed reaction. The observed 3D structural data suggest implications for the enzyme mechanism. It appears that the enzyme-catalyzed cyanohydrin formation is unlikely to proceed via a hemiacetal or hemiketal intermediate covalently attached to the enzyme, despite the observation of such an intermediate for the complex with trichloracetaldehyde. Instead, the data are consistent with a mechanism where the incoming substrate is activated by hydrogen bonding with its carbonyl oxygen to the Ser80 and Thr11 hydroxy groups. A hydrogen cyanide molecule subsequently replaces a water molecule and is deprotonated presumably by the His235 base. Deprotonation is facilitated by the proximity of the positive charge of the Lys236 side chain.

PubMed: 10548044
DOI: 10.1110/ps.8.10.1990

実験手法

X-RAY DIFFRACTION (2 Å)