4DXY

Crystal structures of CYP101D2 Y96A mutant

4DXY の概要

• エントリーDOI: 10.2210/pdb4dxy/pdb
• 関連するPDBエントリー: 3NV5
• 分子名称: Cytochrome P450, PROTOPORPHYRIN IX CONTAINING FE, DI(HYDROXYETHYL)ETHER, ... (4 entities in total)
• 機能のキーワード: cytochrome p450 mutant, haem-dependent, mono-oxygenases, oxidoreductase
• 由来する生物種: Novosphingobium aromaticivorans
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 47946.42

構造登録者

Zhou, W.,Bell, S.G.,Yang, W.,Dale, A.,Wong, L.-L. (登録日: 2012-02-28, 公開日: 2012-08-15, 最終更新日: 2023-11-08)

主引用文献

Bell, S.G.,Yang, W.,Dale, A.,Zhou, W.,Wong, L.-L.
Improving the affinity and activity of CYP101D2 for hydrophobic substrates
Appl.Microbiol.Biotechnol., 2012

PubMed Abstract: CYP101D2 is a cytochrome P450 monooxygenase from Novosphingobium aromaticivorans which is closely related to CYP101A1 (P450cam) from Pseudomonas putida. Both enzymes selectively hydroxylate camphor to 5-exo-hydroxycamphor, and the residues that line the active sites of both enzymes are similar including the pre-eminent Tyr96 residue. However, Met98 and Leu253 in CYP101D2 replace Phe98 and Val247 in CYP101A1, and camphor binding only results in a maximal change in the spin state to 40 % high-spin. Substitutions at Tyr96, Met98 and Leu253 in CYP101D2 reduced both the spin state shift on camphor binding and the camphor oxidation activity. The Tyr96Ala mutant increased the affinity of CYP101D2 for hydrocarbon substrates including adamantane, cyclooctane, hexane and 2-methylpentane. The monooxygenase activity of the Tyr96Ala variant towards alkane substrates was also enhanced compared with the wild-type enzyme. The crystal structure of the substrate-free form of this variant shows the enzyme in an open conformation (PDB: 4DXY), similar to that observed with the wild-type enzyme (PDB: 3NV5), with the side chain of Ala96 pointing away from the heme. Despite this, the binding and activity data suggest that this residue plays an important role in substrate binding, evidencing that the enzyme probably undergoes catalysis in a more closed conformation, similar to those observed in the crystal structures of CYP101A1 (PDB: 2CPP) and CYP101D1 (PDB: 3LXI).

PubMed: 22820521
DOI: 10.1007/s00253-012-4278-7

実験手法

X-RAY DIFFRACTION (2 Å)