Thermal inactivation of reduced ferredoxin (flavodoxin):NADP+ oxidoreductase from Escherichia coli

FEBS Letters

Jarrett, Joseph T¹  Wan, Jason T¹ 

[1] Department of Biochemistry and Biophysics, University of Pennsylvania, 905B Stellar-Chance Laboratories, 422 Curie Boulevard, Philadelphia, PA 19104, USA

关键词: Ferredoxin:NADP reductase;    Flavin-adenine dinucleotide;    Flavoprotein;    Oxidoreductase;    Enzyme stability;    AdoMet;    S-adenosyl-L-methionine;    Bis-Tris Propane;    1;    3-bis[tris(hydroxymethyl)methylamino]propane;    DCIP;    2;    6-dichloroindophenol;    DTT;    dithiothreitol;    Fd;    ferredoxin;    Fld;    flavodoxin;    FNR;    ferredoxin (flavodoxin):NADP+ oxidoreductase;   

DOI  :  10.1016/S0014-5793(02)03349-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Ferredoxin (flavodoxin):NADP⁺ oxidoreductase (FNR) is an essential enzyme that supplies electrons from NADPH to support flavodoxin-dependent enzyme radical generation and enzyme activation. FNR is a monomeric enzyme that contains a non-covalently bound FAD cofactor. We report that reduced FNR from Escherichia coli is subject to inactivation due to unfolding of the protein and dissociation of the FADH₂ cofactor at 37°C. The inactivation rate is temperature-dependent in a manner that parallels the thermal unfolding of the protein and is slowed by binding of ferredoxin or flavodoxin. Understanding factors that minimize inactivation is critical for utilizing FNR as an accessory protein for S-adenosyl-L-methionine-dependent radical enzymes and manipulating FNR as an electron source for biotechnology applications.

【 授权许可】

Unknown