Brzezinski P, Wilson M T
Department of Biochemistry and Biophysics, University of Göteborg and Chalmers University of Technology, Medicinaregatan 9C, S-413 90 Göteborg, Sweden.
Proc Natl Acad Sci U S A. 1997 Jun 10;94(12):6176-9. doi: 10.1073/pnas.94.12.6176.
A new method is presented that makes it possible to inject electrons rapidly into redox-active proteins by means of a short light flash. Reduced carboxymethylated cytochrome c (CmCyt c) with carbon monoxide bound to the heme iron is mixed with the oxidized acceptor protein. Upon rapid photodissociation of CO the apparent redox potential of CmCyt c drops, resulting in electron transfer to the electron acceptor. In this study we have used mitochondrial cytochrome c oxidase as the acceptor protein, but the method also can be used to investigate electron transfer to other proteins that can interact with cytochrome c. In principle, it can be used with any redox protein into which a CO binding site at the heme iron can be engineered.
本文提出了一种新方法,该方法能够通过短光脉冲将电子快速注入氧化还原活性蛋白中。将血红素铁结合有一氧化碳的还原型羧甲基化细胞色素c(CmCyt c)与氧化型受体蛋白混合。一氧化碳快速光解离后,CmCyt c的表观氧化还原电位下降,导致电子转移至电子受体。在本研究中,我们使用线粒体细胞色素c氧化酶作为受体蛋白,但该方法也可用于研究电子向其他能与细胞色素c相互作用的蛋白的转移。原则上,它可用于任何能在血红素铁处设计一氧化碳结合位点的氧化还原蛋白。