细菌ApbC蛋白具有体内功能所需的两种生化活性。
Bacterial ApbC protein has two biochemical activities that are required for in vivo function.
作者信息
Boyd Jeffrey M, Sondelski Jamie L, Downs Diana M
机构信息
Department of Bacteriology, University of Wisconsin, Madison, Wisconsin 53706.
Department of Bacteriology, University of Wisconsin, Madison, Wisconsin 53706.
出版信息
J Biol Chem. 2009 Jan 2;284(1):110-118. doi: 10.1074/jbc.M807003200. Epub 2008 Nov 10.
The ApbC protein has been shown previously to bind and rapidly transfer iron-sulfur ([Fe-S]) clusters to an apoprotein (Boyd, J. M., Pierik, A. J., Netz, D. J., Lill, R., and Downs, D. M. (2008) Biochemistry 47, 8195-8202. This study utilized both in vivo and in vitro assays to examine the function of variant ApbC proteins. The in vivo assays assessed the ability of ApbC proteins to function in pathways with low and high demand for [Fe-S] cluster proteins. Variant ApbC proteins were purified and assayed for the ability to hydrolyze ATP, bind [Fe-S] cluster, and transfer [Fe-S] cluster. This study details the first kinetic analysis of ATP hydrolysis for a member of the ParA subfamily of "deviant" Walker A proteins. Moreover, this study details the first functional analysis of mutant variants of the ever expanding family of ApbC/Nbp35 [Fe-S] cluster biosynthetic proteins. The results herein show that ApbC protein needs ATPase activity and the ability to bind and rapidly transfer [Fe-S] clusters for in vivo function.
先前已证明ApbC蛋白能够结合并迅速将铁硫（[Fe-S]）簇转移至脱辅基蛋白（Boyd, J. M., Pierik, A. J., Netz, D. J., Lill, R., and Downs, D. M. (2008) Biochemistry 47, 8195 - 8202）。本研究利用体内和体外试验来检测变异型ApbC蛋白的功能。体内试验评估了ApbC蛋白在对[Fe-S]簇蛋白需求较低和较高的途径中发挥功能的能力。对变异型ApbC蛋白进行了纯化，并检测其水解ATP、结合[Fe-S]簇以及转移[Fe-S]簇的能力。本研究详细阐述了对“异常”沃克A蛋白ParA亚家族成员ATP水解的首次动力学分析。此外，本研究详细阐述了对不断扩大的ApbC/Nbp35 [Fe-S]簇生物合成蛋白家族突变体变异型的首次功能分析。本文结果表明，ApbC蛋白在体内发挥功能需要ATP酶活性以及结合并迅速转移[Fe-S]簇的能力。
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