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Identification of lysine succinylation as a new post-translational modification

Nature Chemical Biology volume 7, pages 58–63 (2011)Cite this article

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Abstract

Of the 20 ribosomally coded amino acid residues, lysine is the most frequently post-translationally modified, which has important functional and regulatory consequences. Here we report the identification and verification of a previously unreported form of protein post-translational modification (PTM): lysine succinylation. The succinyllysine residue was initially identified by mass spectrometry and protein sequence alignment. The identified succinyllysine peptides derived from in vivo proteins were verified by western blot analysis, in vivo labeling with isotopic succinate, MS/MS and HPLC coelution of their synthetic counterparts. We further show that lysine succinylation is evolutionarily conserved and that this PTM responds to different physiological conditions. Our study also implies that succinyl-CoA might be a cofactor for lysine succinylation. Given the apparent high abundance of lysine succinylation and the significant structural changes induced by this PTM, it is expected that lysine succinylation has important cellular functions.

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Figure 1: Illustration of chemical structures of lysine, acetyllysine, succinyllysine and methylmalonyllysine residues.
Figure 2: Mass spectrometric identification and verification of a lysine-succinylated peptide from isocitrate dehydrogenase.
Figure 3: Verification of lysine succinylation by western blot analysis.
Figure 4: Mass spectrometric identification and verification of a lysine-succinylated peptide from serine hydroxymethyltransferase.
Figure 5: Stimulation of lysine succinylation by sodium succinate and in vivo isotopic succinate labeling.
Figure 6: Sequence alignment and mutagenesis analysis of isocitrate dehydrogenase.

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Acknowledgements

We would like to thank Z. Cheng and Z. Wang for their assistance during the course of this study. This work was supported by US National Institutes of Health grants to Y.Z.

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  1. Zhihong Zhang and Minjia Tan: These authors contributed equally to this work.

Authors and Affiliations

  1. Ben May Department for Cancer Research, The University of Chicago, Chicago, Illinois, USA

    Zhihong Zhang, Minjia Tan, Zhongyu Xie, Lunzhi Dai, Yue Chen & Yingming Zhao

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  1. Zhihong Zhang
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  2. Minjia Tan
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Contributions

Y.Z., M.T. and Z.X. designed the experiments. Y.Z. wrote the paper with the assistance from M.T., Z.Z. and Z.X. Z.Z. contributed to antibody purification and western blots of purified E. coli proteins; M.T. to HPLC-MS and data analysis; Z.X. to western blots, in vivo succinate labeling, mutagenesis, enzymatic assay, circular dichromism experiments, sequence alignment and structural analysis; L.D. to the chemical synthesis; Y.C. to PTMap analysis.

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Correspondence to Yingming Zhao.

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Supplementary information

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Supplementary Methods, Supplementary Figures 1–13, Supplementary Tables 1–3 and Supplementary Data Sets 1 & 2 (PDF 8882 kb)

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Zhang, Z., Tan, M., Xie, Z. et al. Identification of lysine succinylation as a new post-translational modification. Nat Chem Biol 7, 58–63 (2011). https://doi.org/10.1038/nchembio.495

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