Identification of M. tuberculosis Rv3441c and M. smegmatis MSMEG_1556 and essentiality of M. smegmatis MSMEG_1556.
Identification of M. tuberculosis Rv3441c and M. smegmatis MSMEG_1556 and essentiality of M. smegmatis MSMEG_1556.
复制标题
DOI：
10.1371/journal.pone.0042769
复制
发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Ma Y
中科院分区：
文献类型：
--
作者：
Li S;Kang J;Yu W;Zhou Y;Zhang W;Xin Y;Ma Y
分枝杆菌的正常生长依赖于由肽聚糖（PG）、阿拉伯半乳聚糖（AG）和分枝菌酸组成的细胞壁核心的完整性。N-乙酰氨基葡萄糖（GlcNAc）是分枝杆菌细胞壁PG和AG的重要组成部分。UDP-N-乙酰葡萄糖胺（UDP-GlcNAc）作为GlcNAc的糖供体，在原核生物和真核生物中的生物合成途径不同。葡糖胺-6-磷酸转化为葡糖胺-1-磷酸是原核生物所独有的，其由磷酸葡糖胺β（GlmM）催化。生物信息学分析表明，Msm MSMEG_1556和Mtb Rv 3441 c与Ec GlmM同源。本研究在大肠杆菌中表达了可溶性Msm MSMEG_1556蛋白和Mtb Rv 3441 c蛋白。coli BL 21（DE 3），并测定其磷酸葡萄糖胺氧化酶活性。为了进一步研究MSMEG_1556对M.为了获得Mtb Rv 3441 c的条件性MSMEG_1556敲除突变体，我们产生了条件性MSMEG_1556敲除突变体，其具有携带Mtb Rv 3441 c的热敏拯救质粒。由于拯救质粒在42°C下不能补充MSMEG_1556缺陷，因此MSMEG_1556敲除突变体不生长。通过扫描电镜观察到MSMEG_1556基因敲除突变体在30°C至42°C的温度变化后发生了显著的形态学变化。这些结果表明，MSMEG_1556对M.恶臭本研究为GlmM酶作为抗结核药物的潜在靶点提供了证据。
The normal growth of mycobacteria attributes to the integrity of cell wall core which consists of peptidoglycan (PG), arabinogalactan (AG) and mycolic acids. N-acetyl glucosamine (GlcNAc) is an essential component in both PG and AG of mycobacterial cell wall. The biosynthetic pathway for UDP-N-acetylglucosamine (UDP-GlcNAc), as a sugar donor of GlcNAc, is different in prokaryotes and eukaryotes. The conversion of glucosamine-6-phosphate to glucosamine-1-phosphate, which is catalyzed by phosphoglucosamine mutase (GlmM), is unique to prokaryotes. Bioinformatic analysis showed that Msm MSMEG_1556 and Mtb Rv3441c are homologous to Ec GlmM. In this study, soluble Msm MSMEG_1556 protein and Mtb Rv3441c protein were expressed in E. coli BL21(DE3) and their phosphoglucosamine mutase activity were detected. In order to further investigate the essentiality of MSMEG_1556 for the growth of M. smegmatis, we generated a conditional MSMEG_1556 knockout mutant, which harbored thermo-sensitive rescue plasmid carrying Mtb Rv3441c. As the rescue plasmid was unable to complement MSMEG_1556 deficiency at 42°C, MSMEG_1556 knockout mutant did not grow. The dramatic morphological changes of MSMEG_1556 knockout mutant after temperature shift from 30°C to 42°C have been observed by scanning electron microscope. These results demonstrated that MSMEG_1556 is essential for growth of M. smegmatis. This study provided evidence that GlmM enzyme could be as a potential target for developing anti-tuberculosis drugs.
登录
查看更多内容
影响因子：
9.6
作者：
Velayati, Ali Akbar;Masjedi, Mohammad Reza;Hoffner, Sven Eric
通讯作者：
Hoffner, Sven Eric
影响因子：
56.9
作者：
FLEISCHMANN, RD;ADAMS, MD;VENTER, JC
通讯作者：
VENTER, JC
影响因子：
3.2
作者：
Jolly, L;Wu, SW;Tomasz, A
通讯作者：
Tomasz, A
DOI：
10.1046/j.1432-1327.1999.00373.x
发表时间：
1999-05-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
Jolly, L;Ferrari, P;Mengin-Lecreulx, D
通讯作者：
Mengin-Lecreulx, D
影响因子：
14.9
作者：
CORPET, F
通讯作者：
CORPET, F