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Genome Sequence, Proteome Profile, and Identification of a Multiprotein Reductive Dehalogenase Complex in Dehalogenimonas alkenigignens Strain BRE15M
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2020-09-25 , DOI: 10.1021/acs.jproteome.0c00569
Alba Trueba-Santiso 1 , Kenneth Wasmund 2 , Jesica M. Soder-Walz 1 , Ernest Marco-Urrea 1 , Lorenz Adrian 3, 4
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Bacteria of the genus Dehalogenimonas respire with vicinally halogenated alkanes via dihaloelimination. We aimed to describe involved proteins and their supermolecular organization. Metagenomic sequencing of a Dehalogenimonas-containing culture resulted in a 1.65 Mbp draft genome of Dehalogenimonas alkenigignens strain BRE15M. It contained 31 full-length reductive dehalogenase homologous genes (rdhA), but only eight had cognate rdhB gene coding for membrane-anchoring proteins. Shotgun proteomics of cells grown with 1,2-dichloropropane as an electron acceptor identified 1152 proteins representing more than 60% of the total proteome. Ten RdhA proteins were detected, including a DcpA ortholog, which was the strongest expressed RdhA. Blue native gel electrophoresis (BNE) demonstrating maximum activity was localized in a protein complex of 146–242 kDa. Protein mass spectrometry revealed the presence of DcpA, its membrane-anchoring protein DcpB, two hydrogen uptake hydrogenase subunits (HupL and HupS), an iron–sulfur protein (HupX), and subunits of a redox protein with a molybdopterin-binding motif (OmeA and OmeB) in the complex. BNE after protein solubilization with different detergent concentrations revealed no evidence for an interaction between the putative respiratory electron input module (HupLS) and the OmeA/OmeB/HupX module. All detected RdhAs comigrated with the organohalide respiration complex. Based on genomic and proteomic analysis, we propose quinone-independent respiration in Dehalogenimonas.

中文翻译:

基因组序列,蛋白质组概况和多蛋白还原脱卤素酶复合物在Dehalogenimonas alkenigignens菌株BRE15M中的鉴定。

菌属细菌Dehalogenimonas呼吸经由dihaloelimination邻位卤代的烷烃。我们旨在描述涉及的蛋白质及其超分子组织。一个的宏基因组测序Dehalogenimonas含培养导致的1.65 Mbp的基因组草图Dehalogenimonas alkenigignens应变BRE15M。它包含31个全长还原性脱卤素酶同源基因(rdhA),但只有8个具有同源rdhB编码膜固定蛋白的基因。用1,2-二氯丙烷作为电子受体生长的细胞的弹枪蛋白质组学鉴定出1152种蛋白质,占蛋白质组总数的60%以上。检测到十种RdhA蛋白,包括DcpA直向同源物,它是表达最强的RdhA。证明最大活性的蓝色天然凝胶电泳(BNE)位于146-242 kDa的蛋白质复合物中。蛋白质质谱分析表明存在DcpA,其膜锚定蛋白DcpB,两个氢吸收氢化酶亚基(HupL和HupS),铁硫蛋白(HupX)以及具有钼蝶呤结合基序的氧化还原蛋白亚基(OmeA和OmeB)。蛋白质溶解了不同去污剂浓度后的BNE没有显示出假定的呼吸电子输入模块(HupLS)与OmeA / OmeB / HupX模块之间相互作用的证据。所有检测到的RdhA均与有机卤化物呼吸复合物有关。基于基因组和蛋白质组学分析,我们提出了与醌无关的呼吸作用Dehalogenimonas
更新日期:2020-09-25
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