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. 2014 Dec 1;290(6):3308–3332. doi: 10.1074/jbc.M114.611525

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FIGURE 8. — Relative importance of phosphorylatable Ser/Thr and negatively charged Asp/Glu residues in CYP3A4 Glu-258–Ser-259–Glu-262–Asp-263–Thr-264 and Glu-283–Thr-284–Glu-285 surface clusters on its ubiquitination by both gp78 and CHIP complexes. A, ubiquitination of S259A, T264A, S259A/T264A, and E258A/E262A/D263A mutants was compared with that of CYP3A4WT in their native or CuOOH-inactivated form in a system reconstituted with both E2-E3 systems and the presence or absence of PKA/PKC (see under “Experimental Procedures”). B, ubiquitination of E258A/E262A/D263A, E258A/S259A/E262A/D263A/T264A, E283A/E285A, E283A/T284A/E285A, and T264A/S420A/S478A mutants was compared with that of CYP3A4WT in their native or CuOOH-inactivated form, in a system reconstituted with both E2-E3 systems and the presence or absence of PKA/PKC (see under “Experimental Procedures”). The color wheel intensity code is identical to that shown in Fig. 1.