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

被引：15
作者：
Shen, Hongbo [1 ]
Yang, Yanping [1 ]
Wang, Feifei [1 ]
Zhang, Ying [2 ]
Ye, Naihao [3 ]
Xu, Shengfeng [1 ]
Wang, Honghai [1 ]
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
来源：
ACTA BIOCHIMICA ET BIOPHYSICA SINICA | 2009年 / 41卷 / 05期
基金：
中国国家自然科学基金;
关键词：
tryptophan synthase; Mycobacterium tuberculosis; enzyme activity; active site; site-directed mutation; ALPHA-SUBUNIT; ALPHA(2)BETA(2) COMPLEX; SALMONELLA-TYPHIMURIUM; MULTIENZYME COMPLEX; ESCHERICHIA-COLI; MUTAGENESIS; METABOLISM; MORTALITY; CATALYSIS; ALDIMINE;
D O I：
10.1093/abbs/gmp017
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
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 Mg2+ 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.
引用
收藏
页码：379 / 388
页数：10
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