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Proteomic analysis of SUMOylated protein in mammalian cells
Author(s) -
Liu Xiaoyan,
Zhang Fujian,
Zhai Jianjun,
Zhu Haining
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.2_supplement.247
Subject(s) - sumo protein , immunoprecipitation , transfection , microbiology and biotechnology , blot , plasmid , biology , chemistry , cell culture , ubiquitin , gene , genetics
Protein SUMOylation is involved in many cellular activities 1 . The objective of this study is to identify the SUMOylated proteins in mammalian cells. A variety of methods, including cDNA cloning, plasmid construction, cell culture, transfection, western blotting, immunoprecipitation, in‐gel digestion and MS analysis, have been used in the study. We have generated SUMO‐1(T95R) and other truncated SUMO mutants to facilitate the mass spectrometric identification. Seventy four unique proteins were identified using 4 different SUMO1 constructs. Most of these SUMOylated proteins identified are associated with nucleus. Independent immunoblotting assays have validated the SUMOylation status of selected proteins including Ran‐GAP1. In conclusion, a comprehensive study of protein SUMOylation in mammalian cells is accomplished using multiple SUMO‐1 constructs and most SUMOylated proteins identified are related to nucleus.