1. Characterization of aggregates isolated from E
Characterization of aggregates isolated from E. coli cells overexpressing GFP fusion proteins.A, protein aggregates were isolated from cells overexpressing GFP fusion proteins as described under “Experimental Procedures.” The aggregates were analyzed by 1D SDS-PAGE on a 24-cm long 8–16% gradient gel and by 2D gel electrophoresis (B).
Characterization of aggregates isolated from E. coli cells overexpressing GFP fusion proteins.A, protein aggregates were isolated from cells overexpressing GFP fusion proteins as described under “Experimental Procedures.” The aggregates were analyzed by 1D SDS-PAGE on a 24-cm long 8–16% gradient gel and by 2D gel electrophoresis (B). Gels were stained with Coomassie Brilliant Blue R-250. Proteins in indicated bands/spots were identified by MS as described under “Experimental Procedures” (Table I and Supplemental Table 2). C, densitometric analysis of the spot/band intensities in 1D and 2D gels indicated that cytoplasmic chaperones and proteases constituted around 40% of the total protein in aggregates, secretory proteins represented around 25%, the overexpressed membrane protein represented around 20%, and the remainder was cytoplasmic proteins other than chaperones and proteases.
Samuel Wagner et al. Mol Cell Proteomics 2007;6:
© 2007 The American Society for Biochemistry and Molecular Biology