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Incorporation of the prosthetic heme group into cytoplasmatic and membrane proteins

将人工血红素基团掺入细胞质和膜蛋白中

基本信息

批准号：
131891062
负责人：
Professor Dr. Dieter Jahn
金额：
--
依托单位：
Institut für Mikrobiologie
依托单位国家：
德国
项目类别：
Research Units
财政年份：
2009
资助国家：
德国
起止时间：
2008-12-31 至 2015-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/131891062?language=en
关键词：
Incorporation prosthetic heme group into

项目摘要

Heme containing proteins, like enzymes, sensors and electron transporting cytochromes are essential for the metabolism of almost all organisms. Little is known about the transport and insertion of hemes into their cytoplasmic and membrane targets in bacteria. During the last funding period the heme binding protein HemW was identified in Lactococcus lactis. Recombinant anaerobically purified, monomeric Escherichia coli and L. lactis HemW contained a [4Fe-4S] cluster and S-adenosylmethionine (SAM). Upon heme binding the protein forms a dimer in which electrons are transferred from the [4Fe-4S] cluster to heme. The dimer is targeted to the membrane to supply heme in a NADH-dependent reaction to membrane localized electron transfer proteins. Next, we intend to determine the nature of covalent heme binding, the axial heme ligands and the interplay with the [4Fe-4S] cluster and SAM using multiple spectroscopic methods (EPR, Raman resonance, Mössbauer, MCD, ITC). Target enzymes of HemW and potential additional components of the NADHdependent transfer will be identified and characterized. HemW crystal structure determination is intended. HemW related proteins from Bacillus subtilis and human will be investigated. The understanding of the cooperative heme and Moco insertion in model enzymes like nitrate reductase is a long time goal.

含血红素的蛋白质，如酶，传感器和电子传递细胞色素是几乎所有生物体的代谢所必需的。关于血红素在细菌中的转运和插入到它们的细胞质和膜靶点的情况知之甚少。在上一个资助期间，在乳酸乳球菌中鉴定了血红素结合蛋白HemW。重组厌氧纯化的单体大肠杆菌和L. lactis HemW含有[4Fe-4S]簇和S-腺苷甲硫氨酸（SAM）。血红素结合后，蛋白质形成二聚体，其中电子从[4Fe-4S]簇转移到血红素。二聚体靶向膜，以在与膜定位的电子转移蛋白的NADH依赖性反应中提供血红素。接下来，我们打算使用多种光谱方法（EPR，拉曼共振，穆斯堡尔，MCD，ITC）来确定共价血红素结合的性质，轴向血红素配体以及与[4Fe-4S]簇和SAM的相互作用。将鉴定和表征HemW的靶酶和NADH依赖性转移的潜在额外组分。HemW晶体结构的测定。将研究来自枯草芽孢杆菌和人的HemW相关蛋白。理解模型酶如硝酸还原酶中的合作血红素和Moco插入是一个长期的目标。