Recombinant expression and biochemical characterization of Mycobacterium tuberculosis 3Fe-4S ferredoxin Rv1786

被引：0
作者：
Yun Lu
Feng Qiao
Yue Li
Xiao-Hong Sang
Cong-Ran Li
Jian-Dong Jiang
Xin-Yi Yang
Xue-Fu You
机构：
[1] Chinese Academy of Medical Sciences and Peking Union Medical College,Laboratory of Pharmacology/Beijing Key Laboratory of Antimicrobial Agents, Institute of Medicinal Biotechnology
来源：
Applied Microbiology and Biotechnology | 2017年 / 101卷
关键词：
Ferredoxin; Rv1786; Cytochrome P450 mono-oxygenase redox partner; Ferredoxin reductase A; Flavoprotein reductase A;
D O I：
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学科分类号：
摘要：
Ferredoxins are iron-sulfur protein that mediate electron transfer in cytochrome P450 mono-oxygenase (CYP)-related catalytic reactions in a wide variety of organisms. Rv1786 is a putative ferredoxin, encoded by a gene located downstream of the gene encoding CYP143A1 in the Mycobacterium tuberculosis genome. However, the structure and function of Rv1786 have remained unclear. Here, the recombinant Mtb Rv1786 was expressed, purified as a His-tagged form and characterized with [3Fe-4S] clusters as its cofactors using a series of measurements including SDS-PAGE, western blot, UV/Visible, MALDI-TOF/TOF-MS, and electron paramagnetic resonance spectroscopic analysis. Based on the assessments of surface plasmon resonance (SPR) and steady state kinetic assays, Rv1786 was found to be able to couple with both ferredoxin reductase A (FdrA) and flavoprotein reductase A (FprA) as redox partner, but with a stronger binding to FprA and a better coupling activity to FdrA. Preliminary structural and biochemical characterization of Mtb Rv1786 as a redox partner is presented here.
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页码：7201 / 7212
页数：11
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