Heme-dependent autophosphorylation of a heme sensor kinase, ChrS, from Corynebacterium diphtheriae reconstituted in proteoliposomes
Heme-dependent autophosphorylation of a heme sensor kinase, ChrS, from Corynebacterium diphtheriae reconstituted in proteoliposomes
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DOI：
10.1016/j.febslet.2009.06.001
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发表时间：
2009-07-07
期刊：
影响因子：
3.5
通讯作者：
Nakamura, Hiro
中科院分区：
文献类型：
--
作者：
Ito, Yoko;Nakagawa, Shoko;Nakamura, Hiro
白喉棒状杆菌采用双组分信号转导系统的反应调节剂ChrA和传感器激酶ChrS来利用宿主血红素铁。虽然ChrS被预测编码血红素传感器，传感机制仍有待表征。在这份报告中，ChrS表达在大肠杆菌膜溶解和纯化使用癸基麦芽糖苷。ChrS蛋白掺入到蛋白脂质体中，通过ATP催化血红素依赖性自磷酸化。其他金属卟啉和铁没有刺激激酶活性。紫外-可见光谱表明，血红素直接与ChrS相互作用。这是细菌血红素敏感蛋白的第一次功能重建。(C)2009年欧洲生物化学学会联合会。由Elsevier B出版。V.保留所有权利。
Corynebacterium diphteriae employs the response regulator, ChrA, and the sensor kinase, ChrS, of a two-component signal transduction system to utilize host heme iron. Although ChrS is predicted to encode a heme sensor, the sensing mechanism remains to be characterized. In this report, ChrS expressed in Eshcherichia coli membranes was solubilized and purified using decylmaltoside. ChrS protein incorporated into proteoliposomes catalyzed heme-dependent autophosphorylation by ATP. Other metalloporphyrins and iron did not stimulate kinase activity. The UV-Vis spectrum of hemin in the ChrS-proteoliposomes indicated that heme directly interacts with ChrS. This is the first functional reconstitution of a bacterial heme-sensing protein. (C) 2009 Federation of European Biochemical Societies. Published by Elsevier B. V. All rights reserved.