title: Succinylome analysis reveals the involvement of lysine succinylation in metabolism in pathogenic Mycobacterium tuberculosis.
abstract: Mycobacterium tuberculosis (Mtb), the causative agent of human tuberculosis, remains one of the most prevalent human pathogens and a major cause of mortality worldwide. Metabolic network is a central mediator and defining feature of the pathogenicity of Mtb. Increasing evidence suggests that lysine succinylation dynamically regulates enzymes in carbon metabolism in both bacteria and human cells; however, its extent and function in Mtb remain unexplored. Here, we performed a global succinylome analysis of the virulent Mtb strain H37Rv by using high accuracy nano-LC-MS/MS in combination with the enrichment of succinylated peptides from digested cell lysates and subsequent peptide identification. In total, 1545 lysine succinylation sites on 626 proteins were identified in this pathogen. The identified succinylated proteins are involved in various biological processes and a large proportion of the succinylation sites are present on proteins in the central metabolism pathway. Site-specific mutations showed that succinylation is a negative regulatory modification on the enzymatic activity of acetyl-CoA synthetase. Molecular dynamics simulations demonstrated that succinylation affects the conformational stability of acetyl-CoA synthetase, which is critical for its enzymatic activity. Further functional studies showed that CobB, a sirtuin-like deacetylase in Mtb, functions as a desuccinylase of acetyl-CoA synthetase in in vitro assays. Together, our findings reveal widespread roles for lysine succinylation in regulating metabolism and diverse processes in Mtb. Our data provide a rich resource for functional analyses of lysine succinylation and facilitate the dissection of metabolic networks in this life-threatening pathogen.
authors.affiliation: From the ‡Key Laboratory of Algal Biology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China;
authors.forename: Mingkun
authors.initials: M
authors.lastname: Yang
authors.affiliation: From the ‡Key Laboratory of Algal Biology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China;
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: From the ‡Key Laboratory of Algal Biology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China;
authors.forename: Ying
authors.initials: Y
authors.lastname: Chen
authors.affiliation: §Advanced Institute of Translational Medicine, Tongji University, Shanghai 200092, China;
authors.forename: Zhongyi
authors.initials: Z
authors.lastname: Cheng
authors.affiliation: ¶Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China;
authors.forename: Jing
authors.initials: J
authors.lastname: Gu
authors.affiliation: ¶Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China;
authors.forename: Jiaoyu
authors.initials: J
authors.lastname: Deng
authors.affiliation: ‖Key Laboratory of Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China;
authors.forename: Lijun
authors.initials: L
authors.lastname: Bi
authors.affiliation: **Jingjie PTM Biolabs (Hangzhou) Co. Ltd, Hangzhou 310018, China.
authors.forename: Chuangbin
authors.initials: C
authors.lastname: Chen
authors.affiliation: From the ‡Key Laboratory of Algal Biology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China;
authors.forename: Ran
authors.initials: R
authors.lastname: Mo
authors.affiliation: ¶Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China;
authors.forename: Xude
authors.initials: X
authors.lastname: Wang
authors.affiliation: From the ‡Key Laboratory of Algal Biology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; gefeng@ihb.ac.cn.
authors.forename: Feng
authors.initials: F
authors.lastname: Ge
mesh_terms: D000106:Acetate-CoA Ligase / Q000378:metabolism; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D015153:Blotting, Western; D002244:Carbon / Q000494:pharmacology; D017124:Conserved Sequence; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D008239:Lysine / Q000378:metabolism*; D055442:Metabolome / Q000187:drug effects; D055432:Metabolomics; D056004:Molecular Dynamics Simulation; D058977:Molecular Sequence Annotation; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000378:metabolism*; D009243:NAD / Q000378:metabolism; D060066:Protein Interaction Maps / Q000187:drug effects; D021381:Protein Transport / Q000187:drug effects; D015203:Reproducibility of Results; D013386:Succinates / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D013386:Succinates; D009243:NAD; D002244:Carbon; D000106:Acetate-CoA Ligase; D008239:Lysine