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.
Vertegaal
Alfred C O
AC
Wellcome Trust Biocentre, University of Dundee, Dundee DD1 5EH, United Kingdom.
Ogg
Stephen C
SC
Jaffray
Ellis
E
Rodriguez
Manuel S
MS
Hay
Ronald T
RT
Andersen
Jens S
JS
Mann
Matthias
M
Lamond
Angus I
AI
D000951:Antigens, Neoplasm / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics; D002467:Cell Nucleus / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D015870:Gene Expression; D006367:HeLa Cells; D034761:Heterogeneous-Nuclear Ribonucleoprotein Group M / Q000378:metabolism; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D013058:Mass Spectrometry; D008856:Microscopy, Fluorescence; D011499:Protein Processing, Post-Translational; D040901:Proteomics; D034781:Receptors, N-Acetylglucosamine / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D017415:Ribonucleoprotein, U5 Small Nuclear / Q000378:metabolism; D017411:Ribonucleoproteins, Small Nuclear / Q000378:metabolism; D025841:Small Ubiquitin-Related Modifier Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D014357:Trypsin / Q000378:metabolism
D000951:Antigens, Neoplasm; D001426:Bacterial Proteins; C440104:HNRNPM protein, human; D034761:Heterogeneous-Nuclear Ribonucleoprotein Group M; D008164:Luminescent Proteins; D034781:Receptors, N-Acetylglucosamine; D011993:Recombinant Fusion Proteins; D017415:Ribonucleoprotein, U5 Small Nuclear; D017411:Ribonucleoproteins, Small Nuclear; C431323:SART1 protein, human; D025841:Small Ubiquitin-Related Modifier Proteins; C054562:yellow fluorescent protein, Bacteria; D014357:Trypsin