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Originally published In Press as doi:10.1074/mcp.M600212-MCP200 on September 25, 2006.
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Molecular & Cellular Proteomics 5:2298-2310, 2006.
© 2006 by The American Society for Biochemistry and Molecular Biology, Inc.


Research

Distinct and Overlapping Sets of SUMO-1 and SUMO-2 Target Proteins Revealed by Quantitative Proteomics*,S

Alfred C. O. Vertegaal{ddagger},§, Jens S. Andersen, Stephen C. Ogg||, Ronald T. Hay**, Matthias Mann{ddagger}{ddagger} and Angus I. Lamond**,§§

From the {ddagger} Department of Molecular Cell Biology, Leiden University Medical Center, 2300 RC Leiden, The Netherlands, Center for Experimental BioInformatics (CEBI), Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, DK-5230 Odense M, Denmark, || Centre for Molecular Medicine, 61 Biopolis Drive (Proteos), Singapore 138673, Singapore, ** Wellcome Trust Biocentre, University of Dundee, Dundee DD1 5EH, United Kingdom, and {ddagger}{ddagger} Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Am Klopferspitz 18, D-82152 Martinsried, Germany

The small ubiquitin-like modifier (SUMO) family in vertebrates includes three different family members that are conjugated as post-translational modifications to target proteins. SUMO-2 and -3 are nearly identical but differ substantially from SUMO-1. We used quantitative proteomics to investigate the target protein preferences of SUMO-1 and SUMO-2. HeLa cells were established that stably express His6-SUMO-1 or His6-SUMO-2. These cell lines and control HeLa cells were labeled with stable arginine isotopes, and His6-SUMOs were enriched from lysates using immobilized metal affinity chromatography. 53 SUMO-conjugated proteins were identified, including 44 novel SUMO targets. 25 proteins were preferentially conjugated to SUMO-1, 19 were preferentially conjugated to SUMO-2, and nine proteins were conjugated to both SUMO-1 and SUMO-2. SART1 was confirmed by immunoblotting to have both SUMO-1- and SUMO-2-linked forms at similar levels. SUMO-1 and SUMO-2 are thus shown to have distinct and overlapping sets of target proteins, indicating that SUMO-1 and SUMO-2 may have both redundant and non-redundant cellular functions. Interestingly, 14 of the 25 SUMO-1-conjugated proteins contain zinc fingers. Although both SUMO family members play roles in many cellular processes, our data show that sumoylation is strongly associated with transcription because nearly one-third of the identified target proteins are putative transcriptional regulators.


§ To whom correspondence should be addressed: Dept. of Molecular Cell Biology, Leiden University Medical Center, Postal zone S1-P, P O. box 9600, 2300 RC Leiden, The Netherlands. Tel.: 31-71-5269212; Fax: 31-71-5268290; E-mail: vertegaal{at}lumc.nl


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