...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Biochemistry >Intraprotein Electron Transfer in a Two-Domain Construct of Neuronal Nitric Oxide Synthase: The Output State in Nitric Oxide Formation
【24h】

Intraprotein Electron Transfer in a Two-Domain Construct of Neuronal Nitric Oxide Synthase: The Output State in Nitric Oxide Formation

机译：神经元一氧化氮合酶的双结构域构建体中的脑内电子转移：一氧化氮形成中的输出状态

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Intersubunit intraprotein electron transfer (IET) from flavin mononucleotide (FMN) to heme is essential in nitric oxide (NO) synthesis by NO synthase (NOS).Previous crystal structures and functional studies primarily concerned an enzyme conformation,which serves as the input state for reduction of FMN by electrons from NADPH and flavin adenine dinucleotide (FAD) in the reductase domain.To favor the formation of the output state for the subsequent IET from FMN to heme in the oxygenase domain,a novel truncated two-domain oxyFMN construct of rat neuronal NOS (nNOS),in which only the FMN and heme domains were present,was designed and expressed.The kinetics of IET between the FMN and heme domains in the nNOS oxyFMN construct in the presence and absence of added calmodulin (CaM) were directly determined using laser flash photolysis of CO dissociation in comparative studies on partially reduced oxyFMN and single-domain heme oxygenase constructs.The IET rate constant in the presence of CaM (262 s~(-1)) was increased approximately 10-fold compared to that in the absence of CaM (22 s~(-1)).The effect of CaM on interdomain interactions was further evidenced by electron paramagnetic resonance (EPR) spectra.This work provides the first direct evidence of the CaM control of electron transfer (ET) between FMN and heme domains through facilitation of the FMN/heme interactions in the output state.Therefore,CaM controls IET between heme and FMN domains by a conformational gated mechanism.This is essential in coupling ET in the reductase domain in NOS with NO synthesis in the oxygenase domain.
机译：从Flavin单核苷酸（FMN）到血红素的IntersubUnit intaRaprotein电子转移（IET）对于不含合成酶（NoS）的一氧化氮（NO）合成是必不可少的。前晶体结构和功能性研究主要涉及酶构象，其用作输入状态从NADPH和Flavin腺嘌呤二核苷酸（FAD）在还原酶结构域中的电子减少FMN。有利于在氧酶结构域中从FMN到血红素中的后续IET的输出状态形成，一种新型的大鼠截短的双结构型氧杂环构建体设计和表达仅存在FMN和血红色结构域的神经元NOS（NNOS）。在存在和不存在添加的钙调蛋白（CAM）中，NNOS oxyFMN构建体中FMN和血红素结构域之间的动力学是直接的使用激光闪光光解的CO解离在比较研究中，对阳性的氧杂交和单结构域血红素氧酶构建体进行相对研究。在凸轮存在下IET速率恒定（26与凸轮的不存在相比。这项工作通过促进输出状态的FMN / HEME相互作用，提供FMN和血红色域之间的电子转移（ET）的第一直接证据。因此，凸轮通过构象门控通过血红音和FMN域之间的IET控制该机理。该本质在氧酶结构域中没有合成的NoS中的还原酶结构域中是必需的。

著录项

  • 来源
    《Biochemistry》 |2006年第20期|共9页
  • 作者

    Changjian Feng; Gordon Tollin; Michael A.Holliday; Clayton Thomas; John C.Salerno;

  • 作者单位

    College of Pharmacy University of New Mexico Albuquerque New Mexico 87131 Department of Biochemistry and MolecularBiophysics University of Arizona Tucson Arizona 85721 Department of Medicine Duke University and VA Medical Centers Durham North Caroli;

    College of Pharmacy University of New Mexico Albuquerque New Mexico 87131 Department of Biochemistry and MolecularBiophysics University of Arizona Tucson Arizona 85721 Department of Medicine Duke University and VA Medical Centers Durham North Caroli;

    College of Pharmacy University of New Mexico Albuquerque New Mexico 87131 Department of Biochemistry and MolecularBiophysics University of Arizona Tucson Arizona 85721 Department of Medicine Duke University and VA Medical Centers Durham North Caroli;

    College of Pharmacy University of New Mexico Albuquerque New Mexico 87131 Department of Biochemistry and MolecularBiophysics University of Arizona Tucson Arizona 85721 Department of Medicine Duke University and VA Medical Centers Durham North Caroli;

    College of Pharmacy University of New Mexico Albuquerque New Mexico 87131 Department of Biochemistry and MolecularBiophysics University of Arizona Tucson Arizona 85721 Department of Medicine Duke University and VA Medical Centers Durham North Caroli;

  • 收录信息
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 生物化学;
  • 关键词

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号