Differential conjugation of endogenous SUMO-1 and endogenous SUMO-2/3 to target proteins.A and B, SUMO-1 and SUMO-2 proteins were produced in E coli and.

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Differential conjugation of endogenous SUMO-1 and endogenous SUMO-2/3 to target proteins.A and B, SUMO-1 and SUMO-2 proteins were produced in E coli and purified. 5-ng protein samples were subjected to SDS-PAGE, transferred to membranes, and probed using monoclonal antibody 21C7 directed against SUMO-1 (A) and polyclonal antibody AV-SM23-0100 directed against SUMO-2/3 (B). Differential conjugation of endogenous SUMO-1 and endogenous SUMO-2/3 to target proteins.A and B, SUMO-1 and SUMO-2 proteins were produced in E coli and purified. 5-ng protein samples were subjected to SDS-PAGE, transferred to membranes, and probed using monoclonal antibody 21C7 directed against SUMO-1 (A) and polyclonal antibody AV-SM23-0100 directed against SUMO-2/3 (B). C and D, whole cell extracts of HeLa cells were separated by SDS-PAGE, transferred to membranes, and probed using antibodies 21C7 or AV-SM23-0100. Unconjugated SUMO is indicated by an arrow, and SUMO-conjugated RanGAP1 is indicated by an asterisk. The figure is composed of immunoblotting results representing short exposure times (C) and long exposure times (D). Alfred C. O. Vertegaal et al. Mol Cell Proteomics 2006;5:2298-2310 © 2006 The American Society for Biochemistry and Molecular Biology