1JA0

CYPOR-W677X

1JA0 の概要

• エントリーDOI: 10.2210/pdb1ja0/pdb
• 関連するPDBエントリー: 1AMO
• 分子名称: NADPH-Cytochrome P450 Reductase, FLAVIN-ADENINE DINUCLEOTIDE, FLAVIN MONONUCLEOTIDE, ... (5 entities in total)
• 機能のキーワード: nadph-cytochrome p450 reductase, oxidoreductase
• 由来する生物種: Rattus norvegicus (Norway rat)
• 細胞内の位置: Endoplasmic reticulum membrane; Peripheral membrane protein: P00388
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 144328.55

構造登録者

Hubbard, P.A.,Shen, A.L.,Paschke, R.,Kasper, C.B.,Kim, J.J. (登録日: 2001-05-29, 公開日: 2001-08-22, 最終更新日: 2023-08-16)

主引用文献

Hubbard, P.A.,Shen, A.L.,Paschke, R.,Kasper, C.B.,Kim, J.J.
NADPH-cytochrome P450 oxidoreductase. Structural basis for hydride and electron transfer.
J.Biol.Chem., 276:29163-29170, 2001

PubMed Abstract: NADPH-cytochrome P450 oxidoreductase catalyzes transfer of electrons from NADPH, via two flavin cofactors, to various cytochrome P450s. The crystal structure of the rat reductase complexed with NADP(+) has revealed that nicotinamide access to FAD is blocked by an aromatic residue (Trp-677), which stacks against the re-face of the isoalloxazine ring of the flavin. To investigate the nature of interactions between the nicotinamide, FAD, and Trp-677 during the catalytic cycle, three mutant proteins were studied by crystallography. The first mutant, W677X, has the last two C-terminal residues, Trp-677 and Ser-678, removed; the second mutant, W677G, retains the C-terminal serine residue. The third mutant has the following three catalytic residues substituted: S457A, C630A, and D675N. In the W677X and W677G structures, the nicotinamide moiety of NADP(+) lies against the FAD isoalloxazine ring with a tilt of approximately 30 degrees between the planes of the two rings. These results, together with the S457A/C630A/D675N structure, allow us to propose a mechanism for hydride transfer regulated by changes in hydrogen bonding and pi-pi interactions between the isoalloxazine ring and either the nicotinamide ring or Trp-677 indole ring. Superimposition of the mutant and wild-type structures shows significant mobility between the two flavin domains of the enzyme. This, together with the high degree of disorder observed in the FMN domain of all three mutant structures, suggests that conformational changes occur during catalysis.

PubMed: 11371558
DOI: 10.1074/jbc.M101731200

実験手法

X-RAY DIFFRACTION (2.6 Å)