Structural and functional characterization of a mycobacterial methylenetetrahydrofolate reductase utilizing NADH as the exclusive cofactor
文献类型：期刊论文
| 作者 | Li, Jiacong1,2,3; Yang, Mingxia4; Li, Weijia1,5; Lu, Chujie1,5; Feng, Deyu4; Shang, Zhuo6; Wang, Chengyuan4; Lin, Wei1,2,3,5 |
| 刊名 | BIOCHEMICAL JOURNAL
 |
| 出版日期 | 2023-07-01 |
| 卷号 | 480期号:14页码:1129-1146 |
| ISSN号 | 0264-6021 |
| DOI | 10.1042/BCJ20230138 |
| 通讯作者 | Shang, Zhuo(zshang@sdu.edu.cn) ; Wang, Chengyuan(cywang@ips.ac.cn) ; Lin, Wei(weilin@njucm.edu.cn) |
| 英文摘要 | 5,10-Methylenetetraydrofolate reductase (MTHFR) is a key enzyme in folate metabolism. MSMEG_6649, a non-canonical MTHFR from Mycobacterium smegmatis, was previously reported as a monomeric protein lacking the flavin coenzyme. However, the structural basis for its unique flavin-independent catalytic mechanism remains poorly understood. Here, we determined the crystal structures of apo MTHFR MSMEG_6649 and its complex with NADH from M. smegmatis. Structural analysis revealed that the groove formed by the loops 4 and 5 of non-canonical MSMEG_6649 interacting with FAD was significantly larger than that of canonical MTHFR. Meanwhile, the NADH-binding site in MSMEG_6649 is highly similar to the FAD binding site in canonical MTHFR, suggesting that NADH plays the same role (immediate hydride donor for methylenetetraydrofolate) as FAD in the catalytic reaction. Using biochemical analysis, molecular modeling, and sitedirected mutagenesis, the critical residues participating in the binding of NADH and the substrate 5,10-methylenetetrahydrofolate as well as the product 5-methyltetrahydrofolate were identified and validated. Taken together, this work not only provides a good starting point for understanding the potential catalytic mechanism for MSMEG_6649, but also identifies an exploitable target for the development of anti-mycobacterial drugs. |
| WOS关键词 | DEPENDENT 5,10-METHYLENETETRAHYDROFOLATE REDUCTASE ; ESCHERICHIA-COLI ; PURIFICATION ; DIHYDROFOLATE ; ASPARTATE-120 ; TUBERCULOSIS ; CATALYSIS ; ROLES |
| 资助项目 | National Natural Science Foundation of China[81903526] ; National Natural Science Foundation of China[81991523] ; National Natural Science Foundation of China[32270192] ; National Natural Science Foundation of China[82072240] ; State Key Laboratory of Bioreactor Engineering ; State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences[SIMM2205KF-15] ; Chinese Materia Medica First-Class Discipline of Nanjing University of Chinese Medicine[2020YLXK008] ; Fok Ying Tung Education Foundation ; Shanghai Pujiang Program[21PJ1414700] ; Shanghai Municipal Science and Technology Major Project[2019SHZDZX02] |
| WOS研究方向 | Biochemistry & Molecular Biology |
| 语种 | 英语 |
| 出版者 | PORTLAND PRESS LTD |
| WOS记录号 | WOS:001036926800002 |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/306800]  |
| 专题 | 新药研究国家重点实验室 |
| 通讯作者 | Shang, Zhuo; Wang, Chengyuan; Lin, Wei |
| 作者单位 | 1.Nanjing Univ Chinese Med, Sch Med & Holist Integrat Med, Dept Pathogen Biol, Nanjing, Peoples R China 2.East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai, Peoples R China 3.Chinese Acad Sci, Shanghai Inst Mat Med, State Key Lab Drug Res, Shanghai, Peoples R China 4.Chinese Acad Sci, Inst Pasteur Shanghai, Ctr Microbes Dev & Hlth, Shanghai, Peoples R China 5.Nanjing Univ Chinese Med, Sch Pharm, Nanjing, Peoples R China 6.Shandong Univ, Sch Pharmaceut Sci, Jinan, Peoples R China |
| 推荐引用方式 GB/T 7714 | Li, Jiacong,Yang, Mingxia,Li, Weijia,et al. Structural and functional characterization of a mycobacterial methylenetetrahydrofolate reductase utilizing NADH as the exclusive cofactor[J]. BIOCHEMICAL JOURNAL,2023,480(14):1129-1146. |
| APA | Li, Jiacong.,Yang, Mingxia.,Li, Weijia.,Lu, Chujie.,Feng, Deyu.,...&Lin, Wei.(2023).Structural and functional characterization of a mycobacterial methylenetetrahydrofolate reductase utilizing NADH as the exclusive cofactor.BIOCHEMICAL JOURNAL,480(14),1129-1146. |
| MLA | Li, Jiacong,et al."Structural and functional characterization of a mycobacterial methylenetetrahydrofolate reductase utilizing NADH as the exclusive cofactor".BIOCHEMICAL JOURNAL 480.14(2023):1129-1146. |
入库方式： OAI收割
来源：上海药物研究所
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