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. Author manuscript; available in PMC: 2013 Feb 25.
Published in final edited form as: Biochem J. 2011 Jun 15;436(3):581–590. doi: 10.1042/BJ20101446

Figure 3. PolySUMO chain formation is dependent on Asp63 of AtSUMO1.

Figure 3

In vitro SUMOylation assays were performed in the presence of AtSAE2/AtSAE1a, AtSUMO1 or AtSUMO1/D63N and AtSCE1. Reaction mixtures were incubated at 37°C and stopped at the specified time points. Reaction products were resolved by SDS/PAGE and examined by immunoblot analysis with anti-AtSUMO1 (A) or anti-AtSCE1 (B) antibodies. (C) In vitro SUMOylation assays were performed in the presence of AtCAT3Ct as a substrate. Reaction mixtures were incubated at 37°C, aliquots were removed at the specified time points and reaction products were analysed by immunoblot analysis with anti-GST antibodies. The molecular mass in kDa is indicated on the left-hand side.