A proteomic study of SUMO-2 target proteins.
The SUMO family in vertebrates includes at least three distinct proteins (SUMO-1, -2, and -3) that are added as post-translational modifications to target proteins. A considerable number of SUMO-1 target proteins have been identified, but little is known about SUMO-2. A stable HeLa cell line expressing His6-tagged SUMO-2 was established ... and used to label and purify novel endogenous SUMO-2 target proteins. Tagged forms of SUMO-2 were functional and localized predominantly in the nucleus. His6-tagged SUMO-2 conjugates were affinity-purified from nuclear fractions and identified by mass spectrometry. Eight novel potential SUMO-2 target proteins were identified by at least two peptides. Three of these proteins, SART1, heterogeneous nuclear ribonucleoprotein (RNP) M, and the U5 small nuclear RNP 200-kDa helicase, play a role in RNA metabolism. SART1 and heterogeneous nuclear RNP M were both shown to be genuine SUMO targets, confirming the validity of the approach.
Mesh Terms:
Antigens, Neoplasm, Bacterial Proteins, Cell Nucleus, Chromatography, High Pressure Liquid, Gene Expression, Hela Cells, Heterogeneous-Nuclear Ribonucleoprotein Group M, Humans, Luminescent Proteins, Mass Spectrometry, Microscopy, Fluorescence, Protein Processing, Post-Translational, Proteomics, Receptors, N-Acetylglucosamine, Recombinant Fusion Proteins, Ribonucleoprotein, U5 Small Nuclear, Ribonucleoproteins, Small Nuclear, Small Ubiquitin-Related Modifier Proteins, Transfection, Trypsin
Antigens, Neoplasm, Bacterial Proteins, Cell Nucleus, Chromatography, High Pressure Liquid, Gene Expression, Hela Cells, Heterogeneous-Nuclear Ribonucleoprotein Group M, Humans, Luminescent Proteins, Mass Spectrometry, Microscopy, Fluorescence, Protein Processing, Post-Translational, Proteomics, Receptors, N-Acetylglucosamine, Recombinant Fusion Proteins, Ribonucleoprotein, U5 Small Nuclear, Ribonucleoproteins, Small Nuclear, Small Ubiquitin-Related Modifier Proteins, Transfection, Trypsin
J. Biol. Chem.
Date: Aug. 06, 2004
PubMed ID: 15175327
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