1YRC

X-ray Crystal Structure of hydrogenated Cytochrome P450cam

1YRC の概要

• エントリーDOI: 10.2210/pdb1yrc/pdb
• 関連するPDBエントリー: 1DZ4 1DZ6 1DZ8 1DZ9 1YRD 2CPP
• 分子名称: Cytochrome P450-cam, POTASSIUM ION, PROTOPORPHYRIN IX CONTAINING FE, ... (5 entities in total)
• 機能のキーワード: oxidoreductase, mono-oxygenase, heme, ferric
• 由来する生物種: Pseudomonas putida
• 細胞内の位置: Cytoplasm (By similarity): P00183
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 47396.70

構造登録者

Meilleur, F.,Dauvergne, M.-T.,Schlichting, I.,Myles, D.A.A. (登録日: 2005-02-03, 公開日: 2005-02-15, 最終更新日: 2023-10-25)

主引用文献

Meilleur, F.,Dauvergne, M.T.,Schlichting, I.,Myles, D.A.
Production and X-ray crystallographic analysis of fully deuterated cytochrome P450cam.
Acta Crystallogr.,Sect.D, 61:539-544, 2005

PubMed Abstract: Neutron protein crystallography allows H-atom positions to be located in biological structures at the relatively modest resolution of 1.5-2.0 A. A difficulty of this technique arises from the incoherent scattering from hydrogen, which considerably reduces the signal-to-noise ratio of the data. This can be overcome by preparing fully deuterated samples. Efficient protocols for routine and low-cost production of in vivo deuterium-enriched proteins have been developed. Here, the overexpression and crystallization of highly (>99%) deuterium-enriched cytochrome P450cam for neutron analysis is reported. Cytochrome P450cam from Pseudomonas putida catalyses the hydroxylation of camphor from haem-bound molecular O(2) via a mechanism that is thought to involve a proton-shuttle pathway to the active site. Since H atoms cannot be visualized in available X-ray structures, neutron diffraction is being used to determine the protonation states and water structure at the active site of the enzyme. Analysis of both hydrogenated and perdeuterated P450cam showed no significant changes between the X-ray structures determined at 1.4 and 1.7 A, respectively. This work demonstrates that the fully deuterated protein is highly isomorphous with the native (hydrogenated) protein and is appropriate for neutron protein crystallographic analysis.

PubMed: 15858263
DOI: 10.1107/S0907444905003872

実験手法

X-RAY DIFFRACTION (1.4 Å)