Characterization of acetyl-CoA and propionyl-CoA carboxylases encoded by Leptospira interrogans serovar Lai: an initial biochemical study for leptospiral gluconeogenesis via anaplerotic CO_2 assimilation

doi:10.1093/abbs/gms047

	Characterization of acetyl-CoA and propionyl-CoA carboxylases encoded by Leptospira interrogans serovar Lai: an initial biochemical study for leptospiral gluconeogenesis via anaplerotic CO_2 assimilation
	Peng Nanqiu 2; Zhong Yi 2; Zhang Qing 2; Zheng Mingyue 1; Zhao Wei 2; Jiang Hualiang 2; Yang Chen 2; Guo Xiaokui 3; Zhao Guoping 2
刊名	Acta Biochimica et Biophysica Sinica
	2012
卷号	44 期号:8 页码:692
关键词	COENZYME-A CARBOXYLASE STREPTOMYCES-COELICOLOR A3(2) AUTOTROPHIC CARBON FIXATION OUTER-MEMBRANE PROTEIN 3-HYDROXYPROPIONATE CYCLE PATHOGENIC LEPTOSPIRAE SUBSTRATE-SPECIFICITY CITRAMALATE SYNTHASE CRYSTAL-STRUCTURE ESCHERICHIA-COLI acetyl-CoA carboxylase propionyl-CoA carboxylase Leptospira interrogans CO2 assimilation
ISSN号	1672-9145
DOI	10.1093/abbs/gms047
英文摘要	Leptospira interrogans is the causative agent of leptospirosis. The in vitro growth of L. interrogans requires CO2 and a partial 3-hydroxypropionate pathway involving two acyl-CoA carboxylases was suggested by genomic analysis to assimilate CO2. Either set of the candidate genes heterologously co-expressed in Escherichia coli was able to demonstrate both acetyl-CoA carboxylase (ACC) and propionyl-CoA carboxylase (PCC) activities. The tri-subunit holoenzyme (LA_2736-LA_2735 and LA_3803), although failed to be purified, was designated ACC based on its substrate preference toward acetyl-CoA. The partially purified bi-subunit holoenzyme (LA_2432-LA_2433) has a considerably higher activity against propionyl-CoA as the substrate than that of acetyl-CoA, and thus, designated PCC. Native polyacrylamide gel electrophoresis indicated that this PCC has a molecular mass of around 669 kDa, suggesting an (44) quaternary structure and both structural homology modeling and site-directed mutagenesis analysis of its carboxyltransferase subunit (LA_2433) indicated that the A431 residue located at the bottom of the putative substrate binding pocket may play an important role in substrate specificity determination. Both transcriptomic and proteomic data indicated that enzymes involved in the suggested partial 3-hydroxypropionate pathway were expressed in vivo in addition to ACC/PCC and the homologous genes in genomes of other Leptospira species were re-annotated accordingly. However, as the in vitro detected specific activity of ACC in the crude cell extract was too low to account for the growth of the bacterium in EllinghausenMcCulloughJohnsonHarris minimal medium, further systematic analysis is required to unveil the mechanism of gluconeogenesis via anaplerotic CO2 assimilation in Leptospira species.
资助项目	[National Natural Science Foundation of China] ; [National Basic Research Program of China] ; [Research Unit Fund of Li Ka Shing Institute of Health Sciences]
语种	英语
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/286998]
专题	中国科学院上海药物研究所
作者单位	1.中国科学院上海药物研究所 2.中国科学院上海生命科学研究院 3.上海交通大学医学院
推荐引用方式 GB/T 7714	Peng Nanqiu,Zhong Yi,Zhang Qing,et al. Characterization of acetyl-CoA and propionyl-CoA carboxylases encoded by Leptospira interrogans serovar Lai: an initial biochemical study for leptospiral gluconeogenesis via anaplerotic CO_2 assimilation[J]. Acta Biochimica et Biophysica Sinica,2012,44(8):692.
APA	Peng Nanqiu.,Zhong Yi.,Zhang Qing.,Zheng Mingyue.,Zhao Wei.,...&Zhao Guoping.(2012).Characterization of acetyl-CoA and propionyl-CoA carboxylases encoded by Leptospira interrogans serovar Lai: an initial biochemical study for leptospiral gluconeogenesis via anaplerotic CO_2 assimilation.Acta Biochimica et Biophysica Sinica,44(8),692.
MLA	Peng Nanqiu,et al."Characterization of acetyl-CoA and propionyl-CoA carboxylases encoded by Leptospira interrogans serovar Lai: an initial biochemical study for leptospiral gluconeogenesis via anaplerotic CO_2 assimilation".Acta Biochimica et Biophysica Sinica 44.8(2012):692.