title: Proteome-wide Mapping of Endogenous SUMOylation Sites in Mouse Testis.
abstract: SUMOylation is a reversible post-translational modification involved in various critical biological processes. To date, there is limited approach for endogenous wild-type SUMO-modified peptides enrichment and SUMOylation sites identification. In this study, we generated a high-affinity SUMO1 antibody to facilitate the enrichment of endogenous SUMO1-modified peptides from Trypsin/Lys-C protease digestion. Following secondary Glu-C protease digestion, we identified 53 high-confidence SUMO1-modified sites from mouse testis by using high-resolution mass spectrometry. Bioinformatics analyses showed that SUMO1-modified proteins were enriched in transcription regulation and DNA repair. Nab1 was validated to be an authentic SUMOylated protein and Lys479was identified to be the major SUMOylation site. The SUMOylation of Nab1 enhanced its interaction with HDAC2 and maintained its inhibitory effect on EGR1 transcriptional activity. Therefore, we provided a novel approach to investigating endogenous SUMOylation sites in tissue samples.
authors.affiliation: From the ‡State Key Laboratory of Genetic Engineering and Collaborative Innovation Center for Genetics and Development, Institutes of Biomedical Sciences, School of Life Sciences, Fudan University, Shanghai 200032, China.
authors.forename: Lili
authors.initials: L
authors.lastname: Cai
authors.affiliation: From the ‡State Key Laboratory of Genetic Engineering and Collaborative Innovation Center for Genetics and Development, Institutes of Biomedical Sciences, School of Life Sciences, Fudan University, Shanghai 200032, China.|¶Department of Biochemistry and Molecular Cell Biology, Shanghai Key Laboratory for Tumor Microenvironment and Inflammation, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Tu
authors.affiliation: §State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine; National Center for Protein Sciences (The PHOENIX center, Beijing), Beijing 102206, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Song
authors.affiliation: From the ‡State Key Laboratory of Genetic Engineering and Collaborative Innovation Center for Genetics and Development, Institutes of Biomedical Sciences, School of Life Sciences, Fudan University, Shanghai 200032, China.
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Gao
authors.affiliation: §State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine; National Center for Protein Sciences (The PHOENIX center, Beijing), Beijing 102206, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Li
authors.affiliation: From the ‡State Key Laboratory of Genetic Engineering and Collaborative Innovation Center for Genetics and Development, Institutes of Biomedical Sciences, School of Life Sciences, Fudan University, Shanghai 200032, China.
authors.forename: Yunzhi
authors.initials: Y
authors.lastname: Wang
authors.affiliation: From the ‡State Key Laboratory of Genetic Engineering and Collaborative Innovation Center for Genetics and Development, Institutes of Biomedical Sciences, School of Life Sciences, Fudan University, Shanghai 200032, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
authors.affiliation: From the ‡State Key Laboratory of Genetic Engineering and Collaborative Innovation Center for Genetics and Development, Institutes of Biomedical Sciences, School of Life Sciences, Fudan University, Shanghai 200032, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Zhong
authors.affiliation: From the ‡State Key Laboratory of Genetic Engineering and Collaborative Innovation Center for Genetics and Development, Institutes of Biomedical Sciences, School of Life Sciences, Fudan University, Shanghai 200032, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Ge
authors.affiliation: §State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine; National Center for Protein Sciences (The PHOENIX center, Beijing), Beijing 102206, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Qin
authors.affiliation: From the ‡State Key Laboratory of Genetic Engineering and Collaborative Innovation Center for Genetics and Development, Institutes of Biomedical Sciences, School of Life Sciences, Fudan University, Shanghai 200032, China; chend@fudan.edu.cn hefc@nic.bmi.ac.cn.|§State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine; National Center for Protein Sciences (The PHOENIX center, Beijing), Beijing 102206, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Ding
authors.affiliation: From the ‡State Key Laboratory of Genetic Engineering and Collaborative Innovation Center for Genetics and Development, Institutes of Biomedical Sciences, School of Life Sciences, Fudan University, Shanghai 200032, China; chend@fudan.edu.cn hefc@nic.bmi.ac.cn.|§State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine; National Center for Protein Sciences (The PHOENIX center, Beijing), Beijing 102206, China.
authors.forename: Fuchu
authors.initials: F
authors.lastname: He
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000378:metabolism; D019295:Computational Biology; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D008810:Mice, Inbred C57BL; D010455:Peptides / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000378:metabolism*; D015203:Reproducibility of Results; D025842:SUMO-1 Protein / Q000737:chemistry / Q000378:metabolism; D058207:Sumoylation; D013737:Testis / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D000906:Antibodies; D010455:Peptides; D020543:Proteome; D025842:SUMO-1 Protein