3L61

Crystal structure of substrate-free P450cam at 200 mM [K+]

3L61 の概要

• エントリーDOI: 10.2210/pdb3l61/pdb
• 関連するPDBエントリー: 3L62 3L63
• 分子名称: Camphor 5-monooxygenase, PROTOPORPHYRIN IX CONTAINING FE (3 entities in total)
• 機能のキーワード: cytochrome p450, p450cam, camphor, substrate-free, open conformation, metal-binding, monooxygenase, oxidoreductase
• 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: 47173.30

構造登録者

Lee, Y.-T.,Wilson, R.F.,Rupniewski, I.,Goodin, D.B. (登録日: 2009-12-22, 公開日: 2010-04-21, 最終更新日: 2024-04-03)

主引用文献

Lee, Y.T.,Wilson, R.F.,Rupniewski, I.,Goodin, D.B.
P450cam visits an open conformation in the absence of substrate.
Biochemistry, 49:3412-3419, 2010

PubMed Abstract: P450cam from Pseudomonas putida is the best characterized member of the vast family of cytochrome P450s, and it has long been believed to have a more rigid and closed active site relative to other P450s. Here we report X-ray structures of P450cam crystallized in the absence of substrate and at high and low [K(+)]. The camphor-free structures are observed in a distinct open conformation characterized by a water-filled channel created by the retraction of the F and G helices, disorder of the B' helix, and loss of the K(+) binding site. Crystallization in the presence of K(+) alone does not alter the open conformation, while crystallization with camphor alone is sufficient for closure of the channel. Soaking crystals of the open conformation in excess camphor does not promote camphor binding or closure, suggesting resistance to conformational change by the crystal lattice. This open conformation is remarkably similar to that seen upon binding large tethered substrates, showing that it is not the result of a perturbation by the ligand. Redissolved crystals of the open conformation are observed as a mixture of P420 and P450 forms, which is converted to the P450 form upon addition of camphor and K(+). These data reveal that P450cam can dynamically visit an open conformation that allows access to the deeply buried active site without being induced by substrate or ligand.

PubMed: 20297780
DOI: 10.1021/bi100183g

実験手法

X-RAY DIFFRACTION (1.5 Å)