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Characterization of the putative tryptophan synthase beta-subunit from Mycobacterium tuberculosis

文献类型: 外文期刊

作者: Shen, Hongbo 1 ; Yang, Yanping 1 ; Wang, Feifei 1 ; Zhang, Ying 2 ; Ye, Naihao 3 ; Xu, Shengfeng 1 ; Wang, Honghai 1 ;

作者机构: 1.Fudan Univ, Sch Life Sci, State Key Lab Genet Engn, Shanghai 200433, Peoples R China

2.Johns Hopkins Univ, Bloomberg Sch Publ Hlth, Dept Mol Microbiol & Immunol, Baltimore, MD 21205 USA

3.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Qingdao 266071, Peoples R China

关键词: tryptophan synthase;Mycobacterium tuberculosis;enzyme activity;active site;site-directed mutation

期刊名称:ACTA BIOCHIMICA ET BIOPHYSICA SINICA ( 影响因子:3.848; 五年影响因子:3.67 )

ISSN: 1672-9145

年卷期: 2009 年 41 卷 5 期

页码:

收录情况: SCI

摘要: The increasing emergence of drug-resistant tuberculosis (TB) poses a serious threat to the control of this disease. It is in urgent need to develop new TB drugs. Tryptophan biosynthetic pathway plays an important role in the growth and replication of Mycobacterium tuberculosis (Mtb). The beta-subunit of tryptophan synthase (TrpB) catalyzes the last step of the tryptophan biosynthetic pathway, and it might be a potential target for TB drug design. In this study, we overexpressed, purified, and characterized the putative TrpB-encoding gene Rv1612 in Mtb H37Rv. Results showed that Mtb His-TrpB optimal enzymatic activity is at pH 7.8 with 0.15 M Na(+) or 0.18 M Mg(2+) at 37 degrees C. Structure analysis indicated that Mtb TrpB exhibited a typical beta/alpha barrel structure. The amino acid residues believed to interact with the enzyme cofactor pyridoxal-5'-phosphate were predicted by homology modeling and structure alignment. The role of these residues in catalytic activity of the Mtb His-TrpB was confirmed by site-directed mutagenesis. These results provided reassuring structural information for drug design based on TrpB.

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