Fig. 6: Michaelis–Menten analysis of proteasomal degradation.
Degradation of substrate proteins with three different ubiquitin chain modifications by the indicated proteasome types was measured at 30 °C in the presence of 1 mM ATP and monitored by measuring GFP fluorescence. The indicated concentrations of substrate were incubated with 25 nM proteasome, fluorescence was recorded every minute for 120 min, and initial degradation rates were determined by curve fitting. The graphs show the initial rates as a function of substrate concentration. a Ub5(K48)-GFP-35; b Ub9(K48)-GFP-35; c Ub5(K63)-GFP-35 by mutant proteasome. Error bars show standard errors derived from at least three replicate experiments. Source data are provided as a Source Data file.
