1BX1

FERREDOXIN:NADP+ OXIDOREDUCTASE (FERREDOXIN REDUCTASE) MUTANT E312Q

1BX1 の概要

• エントリーDOI: 10.2210/pdb1bx1/pdb
• 分子名称: PROTEIN (FERREDOXIN:NADP+ OXIDOREDUCTASE), PHOSPHATE ION, SULFATE ION, ... (5 entities in total)
• 機能のキーワード: flavoprotein, oxidoreductase
• 由来する生物種: Spinacia oleracea (spinach)
• 細胞内の位置: Plastid, chloroplast stroma: P00455
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 36405.34

構造登録者

Aliverti, A.,Deng, Z.,Ravasi, D.,Piubelli, L.,Karplus, P.A.,Zanetti, G. (登録日: 1998-10-10, 公開日: 1998-10-14, 最終更新日: 2023-08-09)

主引用文献

Aliverti, A.,Deng, Z.,Ravasi, D.,Piubelli, L.,Karplus, P.A.,Zanetti, G.
Probing the function of the invariant glutamyl residue 312 in spinach ferredoxin-NADP+ reductase.
J.Biol.Chem., 273:34008-34015, 1998

PubMed Abstract: Ferredoxin-NADP+ reductase, the prototype of a large family of structurally related flavoenzymes, pairs single electrons carried by ferredoxin I and transfers them as a hydride to NADP+. Four mutants of the enzyme, in which Glu-312 was replaced with Asp, Gln, Leu, and Ala to probe the role of the residue charge, size, and polarity in the enzyme activity, have been heterologously expressed, purified, and characterized through steady-state, rapid kinetic studies, ligand-binding experiments, and three-dimensional structure determination by x-ray crystallography. The E312L mutant was the only one that was almost inactive (approximately 1%), whereas unexpectedly the E312A reductase was 10-100% active with the various acceptors tested. Rapid kinetic absorption spectroscopy studies demonstrated that flavin reduction by NADPH was impaired in the mutants. Furthermore, NADP(H) binding was partially perturbed. These functional and structural studies lead us to conclude that Glu-312 does not fulfil the role of proton donor during catalysis, but it is required for proper binding of the nicotinamide ring of NADP(H). In addition, its charge modulates the two one-electron redox potentials of the flavin to stabilize the semiquinone form.

PubMed: 9852055
DOI: 10.1074/jbc.273.51.34008

実験手法

X-RAY DIFFRACTION (1.9 Å)