Figure 1: Nature Communications

Fig. 1

From: Structural basis of adaptor-mediated protein degradation by the tail-specific PDZ-protease Prc

Fig. 1

NlpI forms a tetrameric complex with Prc and enhances Prc-mediated substrate degradation. a Sedimentation velocity profiles of sNlpI (4.7S), Prc (5.0S), and the sNlpI-Prc complex (9.4S). b SEC-MALS analysis of the sNlpI-Prc complex. UV curve is plotted with fitted molecular weights. c and d Degradation of C- terminal c and N-terminal d 6×His-tagged sMepS (sMepS-6×His-C and N-6×His-sMepS, respectively) by Prc alone or with sNlpI, as monitored by SDS-PAGE assays. sMepS-6×His-C was used in all subsequent assays due to its superior stability. e Thermal shift melting curves (left) and their negative first derivatives (right) of Prc alone and with wild-type NlpI or the binding deficient mutant NlpI-QM (see Fig. 5 below). Melting temperature (T m) values are shown within parentheses. f Degradation of native and 10 mM TCEP-treated denatured lysozyme by Prc alone or with sNlpI. g Degradation of sMepS by the sNlpI-Prc complex inhibited by increasing concentrations of the C-terminal peptide of MepS: NH2-RYNEARRVLSRS-COOH (m). h and i SDS-PAGE assays monitoring degradation of MepS h and denatured lysozyme i by Prc-ΔPDZ without the PDZ domain