The Inhibitory Effect of GlmU Acetyltransferase Inhibitor TPSA on Mycobacterium tuberculosis May Be Affected Due to Its Methylation by Methyltransferase Rv0560c
Mycobacterium tuberculosis bifunctional enzyme GlmU is a novel target for anti-TB drugs and is involved in glycosyl donor UDP-N-acetylglucosamine biosynthesis. Here, we found that TPSA (2-[5-(2-{[4-(2-thienyl)-2-pyrimidinyl]sulfanyl}acetyl)-2-thienyl]acetic acid) was a novel inhibitor for GlmU acety...
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Frontiers Media S.A.
2019-07-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fcimb.2019.00251/full |
| _version_ | 1819174266798604288 |
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| author | Changming Chen Xiuyan Han Qiulong Yan Chao Wang Liqiu Jia Ayaz Taj Lizhe Zhao Yufang Ma Yufang Ma |
| author_facet | Changming Chen Xiuyan Han Qiulong Yan Chao Wang Liqiu Jia Ayaz Taj Lizhe Zhao Yufang Ma Yufang Ma |
| author_sort | Changming Chen |
| collection | DOAJ |
| description | Mycobacterium tuberculosis bifunctional enzyme GlmU is a novel target for anti-TB drugs and is involved in glycosyl donor UDP-N-acetylglucosamine biosynthesis. Here, we found that TPSA (2-[5-(2-{[4-(2-thienyl)-2-pyrimidinyl]sulfanyl}acetyl)-2-thienyl]acetic acid) was a novel inhibitor for GlmU acetyltransferase activity (IC50: 5.3 μM). The interaction sites of GlmU and TPSA by molecular docking were confirmed by site-directed mutagenesis. TPSA showed an inhibitory effect on Mtb H37Ra growth and intracellular H37Ra in macrophage cells (MIC: 66.5 μM). To investigate why TPSA at a higher concentration (66.5 μM) was able to inhibit H37Ra growth, proteome and transcriptome of H37Ra treated with TPSA were analyzed. The expression of two methyltransferases MRA_0565 (Rv0558) and MRA_0567 (Rv0560c) were markedly increased. TPSA was pre-incubated with purified Rv0558 and Rv0560c in the presence of S-adenosylmethionine (methyl donor) respectively, resulting in its decreased inhibitory effect of GlmU on acetyltransferase activity. The inhibition of TPSA on growth of H37Ra with overexpressed Rv0558 and Rv0560c was reduced. These implied that methyltransferases could modify TPSA. The methylation of TPSA catalyzed by Rv0560c was subsequently confirmed by LC-MS. Therefore, TPSA as a GlmU acetyltransferase activity inhibitor may offer a structural basis for new anti-tuberculosis drugs. TPSA needs to be modified further by some groups to prevent its methylation by methyltransferases. |
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| language | English |
| last_indexed | 2024-12-22T20:36:15Z |
| publishDate | 2019-07-01 |
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| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-9fb555eb3c984b5f9da284c6d22f702a2022-12-21T18:13:27ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882019-07-01910.3389/fcimb.2019.00251459446The Inhibitory Effect of GlmU Acetyltransferase Inhibitor TPSA on Mycobacterium tuberculosis May Be Affected Due to Its Methylation by Methyltransferase Rv0560cChangming Chen0Xiuyan Han1Qiulong Yan2Chao Wang3Liqiu Jia4Ayaz Taj5Lizhe Zhao6Yufang Ma7Yufang Ma8Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Dalian Medical University, Dalian, ChinaDepartment of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Dalian Medical University, Dalian, ChinaDepartment of Microbiology, College of Basic Medical Sciences, Dalian Medical University, Dalian, ChinaCollege of Pharmacy, Dalian Medical University, Dalian, ChinaDepartment of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Dalian Medical University, Dalian, ChinaDepartment of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Dalian Medical University, Dalian, ChinaDepartment of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Dalian Medical University, Dalian, ChinaDepartment of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Dalian Medical University, Dalian, ChinaDepartment of Microbiology, College of Basic Medical Sciences, Dalian Medical University, Dalian, ChinaMycobacterium tuberculosis bifunctional enzyme GlmU is a novel target for anti-TB drugs and is involved in glycosyl donor UDP-N-acetylglucosamine biosynthesis. Here, we found that TPSA (2-[5-(2-{[4-(2-thienyl)-2-pyrimidinyl]sulfanyl}acetyl)-2-thienyl]acetic acid) was a novel inhibitor for GlmU acetyltransferase activity (IC50: 5.3 μM). The interaction sites of GlmU and TPSA by molecular docking were confirmed by site-directed mutagenesis. TPSA showed an inhibitory effect on Mtb H37Ra growth and intracellular H37Ra in macrophage cells (MIC: 66.5 μM). To investigate why TPSA at a higher concentration (66.5 μM) was able to inhibit H37Ra growth, proteome and transcriptome of H37Ra treated with TPSA were analyzed. The expression of two methyltransferases MRA_0565 (Rv0558) and MRA_0567 (Rv0560c) were markedly increased. TPSA was pre-incubated with purified Rv0558 and Rv0560c in the presence of S-adenosylmethionine (methyl donor) respectively, resulting in its decreased inhibitory effect of GlmU on acetyltransferase activity. The inhibition of TPSA on growth of H37Ra with overexpressed Rv0558 and Rv0560c was reduced. These implied that methyltransferases could modify TPSA. The methylation of TPSA catalyzed by Rv0560c was subsequently confirmed by LC-MS. Therefore, TPSA as a GlmU acetyltransferase activity inhibitor may offer a structural basis for new anti-tuberculosis drugs. TPSA needs to be modified further by some groups to prevent its methylation by methyltransferases.https://www.frontiersin.org/article/10.3389/fcimb.2019.00251/fullMycobacteria tuberculosisUDP-GlcNAcGlmU acetyltransferaseinhibitormethyltransferases |
| spellingShingle | Changming Chen Xiuyan Han Qiulong Yan Chao Wang Liqiu Jia Ayaz Taj Lizhe Zhao Yufang Ma Yufang Ma The Inhibitory Effect of GlmU Acetyltransferase Inhibitor TPSA on Mycobacterium tuberculosis May Be Affected Due to Its Methylation by Methyltransferase Rv0560c Frontiers in Cellular and Infection Microbiology Mycobacteria tuberculosis UDP-GlcNAc GlmU acetyltransferase inhibitor methyltransferases |
| title | The Inhibitory Effect of GlmU Acetyltransferase Inhibitor TPSA on Mycobacterium tuberculosis May Be Affected Due to Its Methylation by Methyltransferase Rv0560c |
| title_full | The Inhibitory Effect of GlmU Acetyltransferase Inhibitor TPSA on Mycobacterium tuberculosis May Be Affected Due to Its Methylation by Methyltransferase Rv0560c |
| title_fullStr | The Inhibitory Effect of GlmU Acetyltransferase Inhibitor TPSA on Mycobacterium tuberculosis May Be Affected Due to Its Methylation by Methyltransferase Rv0560c |
| title_full_unstemmed | The Inhibitory Effect of GlmU Acetyltransferase Inhibitor TPSA on Mycobacterium tuberculosis May Be Affected Due to Its Methylation by Methyltransferase Rv0560c |
| title_short | The Inhibitory Effect of GlmU Acetyltransferase Inhibitor TPSA on Mycobacterium tuberculosis May Be Affected Due to Its Methylation by Methyltransferase Rv0560c |
| title_sort | inhibitory effect of glmu acetyltransferase inhibitor tpsa on mycobacterium tuberculosis may be affected due to its methylation by methyltransferase rv0560c |
| topic | Mycobacteria tuberculosis UDP-GlcNAc GlmU acetyltransferase inhibitor methyltransferases |
| url | https://www.frontiersin.org/article/10.3389/fcimb.2019.00251/full |
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