Mapping the cytochrome c 553 interacting site using 1H and 15N NMR

FEBS Letters

Guerlesquin, Françoise¹  Morelli, Xavier¹ 

[1] Unité de Bioénergétique et Ingénierie des Protéines, IBSM-CNRS, Marseille, France

关键词: Cytochrome c 553;    Ferredoxin;    1H and 15N NMR;    Protein-protein interaction;    NMR;    nuclear magnetic resonance;    HSQC;    heteronuclear single quantum coherence;    DdN;    Desulfovibrio desulfuricans Norway;    DvH;    Desulfovibrio vulgaris Hildenborough;   

DOI  :  10.1016/S0014-5793(99)01299-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Cytochrome c ₅₅₃ is the electron transfer partner of formate dehydrogenase and of [Fe]-hydrogenase, two metalloenzymes essential in the metabolism of sulfate reducing bacteria. These two enzymes contain a ‘ferredoxin-like’ domain which presents 30% identity with Desulfovibrio desulfuricans Norway ferredoxin I. This was chosen as a model for the ‘ferredoxin-like’ domain involved in the electron transfer reaction with cytochrome c ₅₅₃. 1D NMR titration of complex formation gave us the stoichiometry (1:1) and the dissociation constant of the complex (K _d ∼3×10⁻⁶ M). 2D heteronuclear NMR experiments were performed to analyze the ¹H and ¹⁵N chemical shift variations that are induced by the protein-protein recognition. This is the first mapping of the interaction site on a c-type cytochrome, using heteronuclear NMR.

【 授权许可】

Unknown