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首页> 外文期刊>RSC Advances >Electron transfer amongst flavo- and hemo-proteins: diffusible species effect the relay processes, not protein-protein binding
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Electron transfer amongst flavo- and hemo-proteins: diffusible species effect the relay processes, not protein-protein binding

机译：电子转移在黄色和血液蛋白中：扩散物种影响继电器工艺，而不是蛋白质蛋白质结合

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摘要

Hitherto, electron transfer (ET) between redox proteins has been deemed to occur via donor-acceptor binding, and diffusible reactive species are considered as deleterious side-products in such systems. Herein, ET from cytochrome P450 reductase (CPR, an animal membrane flavoprotein) and horseradish peroxidase (HRP, a plant hemoprotein) to cytochrome c (Cyt c, a soluble animal hemoprotein) was probed under diverse conditions, using standard assays. ET in the CPR-Cyt c system was critically inhibited by cyanide and sub-equivalent levels of polar one-electron cyclers like copper ions, vitamin C/Trolox and superoxide dismutase. In the presence of lipids, inhibition was also afforded by amphipathic molecules vitamin E, palmitoyl-vitamin C and the membrane hemoprotein, cytochrome b(5). Such nonspecific inhibition (by diverse agents in both aqueous and lipid phases) indicated that electron transfer/relay was effected by small diffusible agents, whose lifetimes are shortened by the diverse radical scavengers. When CPR was retained in a dialysis membrane and Cyt c presented outside in free solution, ET was still observed. Further, HRP (taken at nM levels) catalyzed oxidation of a phenolic substrate was significantly inhibited upon the incorporation of sub-nM levels of Cyt c. The findings imply that CPR-Cyt c or HRP-Cyt c binding is not crucial for ET. Further, fundamental quantitative arguments (based on diffusion/collision) challenge the erstwhile protein-protein binding-assisted ET hypothesis. It is proven beyond reasonable doubt that mobile and diffusible electron carriers (ions and radicals) serve as "redox-relay agents" in the biological ET models/setup studied.
机译：迄今为止，氧化还原蛋白之间的电子转移（ET）被认为是通过供体 - 受体结合发生的，并且在这种系统中被认为是有害的反应物质被认为是有害的副产物。在本文中，在不同的条件下，探测了在不同条件下探测到细胞色素P450还原酶（CPR，动物膜黄蛋白）和辣根过氧化物酶（HRP，植物杂液），使用标准测定。在CPR-CYT C系统中，通过氰化物和亚等当量水平的极性单电子循环助剂，如铜离子，维生素C / TROLOX和超氧化物歧化酶。在存在脂质存在下，还通过两亲分子维生素E，Palmitoyl-ViTamin C和膜杂液，细胞色素B（5）提供抑制。这种非特异性抑制（通过水性和脂质相中的多种试剂）表明电子转移/继电器由小扩散剂进行，其寿命由不同的自由基清除剂缩短。当CPR保留在透析膜中，并且在游离溶液中呈现外部的Cyt C时，ET仍然观察到。此外，在掺入亚NM水平的Cyt C时，HRP催化酚基质的氧化显着抑制。研究结果暗示CPR-CYT C或HRP-CYT C结合对ET至关重要。此外，基本的定量争论（基于扩散/碰撞）挑战ETSHILE蛋白质结合辅助等假设。超越合理怀疑，移动和扩散的电子载体（离子和基团）作为所研究的生物ET模型/设置中的“氧化还原剂”作为“氧化还原继电器”。

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  • 来源
    《RSC Advances》 |2016年第29期|共9页
  • 作者

    Manoj Kelath Murali; Gade Sudeep K.; Venkatachalam Avanthika; Gideon Daniel A.;

  • 作者单位

    Satyamjayatu Sci &

    Eth Fdn Shoranur 2 Kulappully 679122 Kerala India;

    VIT Univ Sch Biosci &

    Technol Hemoprot Lab Vellore 632014 Tamil Nadu India;

    PSG Inst Adv Studies REDOx Lab Caimbatare Tamil Nadu India;

    Indian Inst Sci CIDR MCBL Bengaluru 560012 Karnataka India;

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