Preferential conjugation of proteins to SUMO-1 or SUMO-2

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Preferential conjugation of proteins to SUMO-1 or SUMO-2 Preferential conjugation of proteins to SUMO-1 or SUMO-2.A, His6-SUMO conjugates were purified from HeLaHis6-SUMO-1 nuclei and HeLaHis6-SUMO-2 nuclei. Preferential conjugation of proteins to SUMO-1 or SUMO-2.A, His6-SUMO conjugates were purified from HeLaHis6-SUMO-1 nuclei and HeLaHis6-SUMO-2 nuclei. Control purifications from HeLa nuclei were included in the experiment. Proteins were size-separated by SDS-PAGE, transferred to membranes, and probed using antibodies directed against endogenous SUMO target proteins. RanGAP1 is preferentially conjugated to SUMO-1, Sp100 is conjugated to SUMO-2, and SART1 is conjugated to both SUMOs at similar levels. B, preferential conjugation of endogenous target proteins to endogenous SUMO-1 or endogenous SUMO-2/3. The SUMO-1 target protein RanGAP1 and the SUMO-2 target protein Sp100 were immunoprecipitated from HeLa cell lysates, transferred to a membrane, and probed using antibody 21C7 directed against SUMO-1 or antibody AV-SM23-0100 directed against SUMO-2/3. Alfred C. O. Vertegaal et al. Mol Cell Proteomics 2006;5:2298-2310 © 2006 The American Society for Biochemistry and Molecular Biology