Characterization of Mycobacterium tuberculosis nicotinamidase/pyrazinamidase

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	Characterization of Mycobacterium tuberculosis nicotinamidase/pyrazinamidase
	Zhang, H; Deng, JY; Bi, LJ; Zhou, YF; Zhang, ZP; Zhang, CG; Zhang, Y; Zhang, XE; Zhang, XE, Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, Wuchang Dist, Wuhan 430071, Peoples R China
刊名	FEBS JOURNAL
	2008
卷号	275 期号:4 页码:753-762
关键词	Mycobacterium Tuberculosis Nicotinamidase Pnca Pyrazinamidase Site-directed Mutation
ISSN号	1742-464X
英文摘要	The nicotinamidase/pyrazinamidase (PncA) of Mycobacterium tuberculosis is involved in the activation of the important front-line antituberculosis drug pyrazinamide by converting it into the active form, pyrazinoic acid. Mutations in the pncA gene cause pyrazinamide resistance in M. tuberculosis. The properties of M. tuberculosis PncA were characterized in this study. The enzyme was found to be a 20.89 kDa monomeric protein. The optimal pH and temperature of enzymatic activity were pH 7.0 and 40 degrees C, respectively. Inductively coupled plasma-optical emission spectrometry revealed that the enzyme was an Mn2+/Fe2+-containing protein with a molar ratio of [Mn2+] to [Fe2+] of 1 : 1; furthermore, the external addition of either type of metal ion had no apparent effect on the wild-type enzymatic activity. The activity of the purified enzyme was determined by HPLC, and it was shown that it possessed similar pyrazinamidase and nicotinamidase activity, by contrast with previous reports. Nine PncA mutants were generated by site-directed mutagenesis. Determination of the enzymatic activity and metal ion content suggested that Asp8, Lys96 and Cys138 were key residues for catalysis, and Asp49, His51, His57 and His71 were essential for metal ion binding. Our data show that M. tuberculosis PncA may bind metal ions in a manner different from that observed in the case of Pyrococcus horikoshii PncA.
学科主题	Biochemistry & Molecular Biology
语种	英语
WOS记录号	WOS:000252712000016
公开日期	2011-09-23
内容类型	期刊论文
源URL	[http://210.72.129.5/handle/321005/54539]
专题	沈阳应用生态研究所_沈阳应用生态研究所
通讯作者	Zhang, XE, Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, Wuchang Dist, Wuhan 430071, Peoples R China
推荐引用方式 GB/T 7714	Zhang, H,Deng, JY,Bi, LJ,et al. Characterization of Mycobacterium tuberculosis nicotinamidase/pyrazinamidase[J]. FEBS JOURNAL,2008,275(4):753-762.
APA	Zhang, H.,Deng, JY.,Bi, LJ.,Zhou, YF.,Zhang, ZP.,...&Zhang, XE, Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, Wuchang Dist, Wuhan 430071, Peoples R China.(2008).Characterization of Mycobacterium tuberculosis nicotinamidase/pyrazinamidase.FEBS JOURNAL,275(4),753-762.
MLA	Zhang, H,et al."Characterization of Mycobacterium tuberculosis nicotinamidase/pyrazinamidase".FEBS JOURNAL 275.4(2008):753-762.