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首页> 外文期刊>Biochemistry >Redox Potential and Equilibria in the Reductive Half-Reaction of Vibrio harveyi NADPH-FMN Oxidoreductase
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Redox Potential and Equilibria in the Reductive Half-Reaction of Vibrio harveyi NADPH-FMN Oxidoreductase

机译：氧化还原潜力和均衡在哈维群岛NADPH-FMN氧化还原酶的还原半反应中

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摘要

Vibrio harveyi NADPH:FMN oxidoreductase P (FRPvh) is a homodimeric enzyme having a bound FMN per enzyme monomer.The bound FMN functions as a cofactor of FRP_(Vh) in transferring reducing equivalents from NADPH to a flavin substrate in the absence of V.harveyi luciferase but as a substrate for FRP_(Vh) in the luciferase-coupled bioluminescent reaction.As part of an integral plan to elucidate the regulation of functional coupling between FRPvh and luciferase,this study was carried out to characterize the equilibrium bindings,reductive potential,and the reversibility of the reduction of the bound FMN in the reductive half-reaction of FRPyh- Results indicate that,in addition to NADPH binding,NADP~+ also bound to FRPvh in either the oxidized (Kd 180 muM) or reduced (K_d 230 muM) form.By titrations with NADP+ and NADPH and by an isotope exchange experiment,the reduction of the bound FMN by NADPH was found to be readily reversible (K_(eq) = 0.8).Hence,the reduction of FRPvh-bound FMN is not the committed step in coupling the NADPH oxidation to bioluminescence.To our knowledge,such an aspect of flavin reductase catalysis has only been clearly established for FRPvh- Although the reductive potentials and some other properties of a R203 A variant of FRPvh and an NADH/NADPH-utilizing flavin reductase from Vibrio fischeri are quite similar to that of the wild-type FRPvh,the reversal of the reduction of bound FMN was not detected for either of these two enzymes.
机译：vibrio harveyi nadph：fmn氧化还原酶p（frpvh）是每种酶单体的结合FMN的同源二聚体酶。结合的FMN作为FRP_（VH）的辅因子，在不存在V的情况下从NADPH转移到黄蛋白基质中的降低等同物。 Harveyi Luciferase但作为荧光素酶偶联的生物发光反应中的FRP_（VH）的基材。为了阐明FRPVH和荧光素酶之间的功能偶联调节的一部分的一部分，该研究进行了均衡结合，还原潜力并且，在FrPyH的还原半反应中，减少结合FMN的可逆性表明，除了NADPH结合外，NADP〜+还与氧化（KD 180妈妈）或降低的FRPVH结合（K_D 230毫米）形式。用NADP +和NADPH的滴定和同位素交换实验，发现NADPH的结合FMN的减少是容易可逆的（K_（EQ）= 0.8）。，降低FRPVH绑定的FMN不是t.他致力于偶联NADPH氧化对生物发光。对于我们的知识，SPRAVIN还原酶催化的这种方面仅用于FRPVH-虽然R203 R203的R203变体和NADH / NADPH的变种 - 从vibrio fischeri的utilized flavin还原酶与野生型frpvh的化合物相似，对这两种酶中的任何一种没有检测到结合的fmn减少的反转。

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  • 来源
    《Biochemistry》 |2005年第1期|共7页
  • 作者

    Benfang Lei; He Wang; Yimin Yu; Shiao-Chun Tu;

  • 作者单位

    Department of Biology and Biochemistry University of Houston;

    Houston Texas 77204-5001;

    Department of Biology and Biochemistry University of Houston;

    Houston Texas 77204-5001;

    Department of Biology and Biochemistry University of Houston;

    Houston Texas 77204-5001;

    Department of Biology and Biochemistry University of Houston;

    Houston Texas 77204-5001;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 生物化学;
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