作者
Luiz Pedro S de Carvalho, Hong Zhao, Caitlyn E Dickinson, Nancy M Arango, Christopher D Lima, Steven M Fischer, Ouathek Ouerfelli, Carl Nathan, Kyu Y Rhee
发表日期
2010/4/23
期刊
Chemistry & biology
卷号
17
期号
4
页码范围
323-332
出版商
Elsevier
简介
Activity based metabolomic profiling (ABMP) allows unbiased discovery of enzymatic activities encoded by genes of unknown function, and applies liquid-chromatography mass spectrometry (LC-MS) to analyze the impact of a recombinant enzyme on the homologous cellular extract as a physiologic library of potential substrates and products. The Mycobacterium tuberculosis protein Rv1248c was incompletely characterized as a thiamine diphosphate-dependent α-ketoglutarate decarboxylase. Here, recombinant Rv1248c catalyzed consumption of α-ketoglutarate in a mycobacterial small molecule extract with matched production of 5-hydroxylevulinate (HLA) in a reaction predicted to require glyoxylate. As confirmed using pure substrates by LC-MS, 1H-NMR, chemical trapping, and intracellular metabolite profiling, Rv1248c catalyzes C-C bond formation between the activated aldehyde of α–ketoglutarate and the …
学术搜索中的文章
LPS de Carvalho, H Zhao, CE Dickinson, NM Arango… - Chemistry & biology, 2010