5X7E

Crystal structure of vitamin D hydroxylase cytochrome P450 105A1 (R84A mutant) in complex with 1,25-dihydroxyvitamin D2

5X7E の概要

• エントリーDOI: 10.2210/pdb5x7e/pdb
• 分子名称: Vitamin D3 dihydroxylase, PROTOPORPHYRIN IX CONTAINING FE, (1R,3S,5Z)-5-[(2E)-2-[(1R,3aS,7aR)-1-[(E,2R,5S)-5,6-dimethyl-6-oxidanyl-hept-3-en-2-yl]-7a-methyl-2,3,3a,5,6,7-hexahydr o-1H-inden-4-ylidene]ethylidene]-4-methylidene-cyclohexane-1,3-diol, ... (4 entities in total)
• 機能のキーワード: metal-binding, monooxygenase, oxidoreductase
• 由来する生物種: Streptomyces griseolus
• 細胞内の位置: Cytoplasm : P18326
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 46036.88

構造登録者

Hayashi, K.,Yasuda, K.,Shiro, Y.,Sugimoto, H.,Sakaki, T. (登録日: 2017-02-25, 公開日: 2017-05-10, 最終更新日: 2023-11-22)

主引用文献

Yasuda, K.,Yogo, Y.,Sugimoto, H.,Mano, H.,Takita, T.,Ohta, M.,Kamakura, M.,Ikushiro, S.,Yasukawa, K.,Shiro, Y.,Sakaki, T.
Production of an active form of vitamin D2 by genetically engineered CYP105A1
Biochem. Biophys. Res. Commun., 486:336-341, 2017

PubMed Abstract: Our previous studies revealed that CYP105A1 can convert vitamin D (VD3) to its active form, 1α,25-dihydroxyvitamin D (1,25D3). Site-directed mutagenesis of CYP105A1 based on its crystal structure dramatically enhanced its activity; the activity of double variants R73A/R84A and R73A/R84V was more than 100-fold higher than that of the wild type of CYP105A1. In contrast, these variants had a low ability to convert vitamin D (VD2) to 1α,25-dihydroxyvitamin D (1,25D2), whereas they catalyzed the sequential hydroxylation at positions C25 and C26 to produce 25,26D2. A comparison of the docking models of 25D2 and 25D3 into the substrate-binding pocket of R73A/R84A suggests that the side chain of the Met239 inhibits the binding of 25D2 for 1α-hydroxylation. Therefore, the Met239 residue of R73A/R84A was substituted for Ala. As expected, the triple variant R73A/R84A/M239A showed a 22-fold higher 1α-hydroxylation activity towards 25D2. To the best of our knowledge, this is the first report on the generation of microbial cytochrome P450 that converts VD2 to 1,25D2 via 25D2.

PubMed: 28302483
DOI: 10.1016/j.bbrc.2017.03.040

実験手法

X-RAY DIFFRACTION (1.9 Å)