The non‐equivalence of binding sites of coenzyme quinone and rotenone in mitochondrial NADH‐CoQ reductase

FEBS Letters

Ahmed, Ishak¹  Krishnamoorthy, G.¹ 

[1] Chemical Physics Group, Tata Institute of Fundamental Research, Homi Bhabha Road, Bombay 400 005, India

关键词: NADH-CoQ reductase;    Erythrosine-5′-iodoacetamide;    Fluorescence probe;    Temperature jump;    Rotenone;    ER;    erythrosine-5′-iodoacetamide;    DB;    2;    3-dimethoxy-5-methyl-6-decyl-1;    4-benzoquinone;    PDB;    2;    3-dimethoxy-5-methyl-1;    4-benzoquinone;    T-jump;    temperature jump;    MES;    2-[N-morpholino]ethanesulphonic acid;    MOPS;    3-[N-morpholino]-propanesulphonic acid;    CAPSO;    3-[cyclohexylamino]-2-hydroxy-1-propanesulphonic acid;    Tris;    Tris(hydroxymethyl)aminomethane;   

DOI  :  10.1016/0014-5793(92)80862-B

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The fluorescent probe erythrosine 5′-iodoacetamide (ER) binds to mitochondrial NADH-CoQ reductase (Complex-I) accompanied by an enhancement of the fluorescence intensity. The binding of the CoQ analogue, 2,3-dimethoxy-5-methyl-6-decyl-1,4-benzoquinone (DB), decreased the fluorescence intensity of the ER:Complex-I system. The ‘site 1’ inhibitor rotenone did not decrease the fluorescence intensity showing the non-identical nature of the binding sites of DB and rotenone. Also, the reduced form of DB did not decrease the fluorescence intensity. The decrease of the fluorescence intensity by DB was shown to be due to the removal of bound ER by DB. The rapid kinetics of ER binding was studied by temperature-jump relaxation. While DB caused complete elimination of the relaxation process in the ER:Complex-I system, rotenone caused only a decrease in the relaxation rate, suggesting conformational change. The relaxation rate showed a pH dependence with a maximum around pH 7.5.

【 授权许可】

Unknown