Figure 4. The Msm Mpa/Opengate proteasome complex does not degrade Pup-GGQ.

A. Overlay of E. coli ¹H{¹⁵N}-HSQC spectra showing [U- ¹⁵N] Pup-GGQ after 4 h of over-expression (red), and after 4 h of over-expression following 16 h of Msm Mpa/Opengate proteasome complex over-expression (black). B. Overlay of E. coli ¹H{¹⁵N}-HSQC spectra of [U- ¹⁵N] Pup-GGQ after 4 h of over-expression (red) followed by 16 h of Msm Mpa/Opengate proteasome complex over-expression. C. Sequence of Pup-GGQ showing the greatest intensity changes upon complex formation in red. D & E. Western blots showing the effect of Msm Mpa and the Opengate proteasome CP over-expression on Pup-GGQ induction, probed with anti-His-tag HRP-conjugated antibody. D. Pup-GGQ was over-expressed first for 6 h, followed by over-expression of Msm Mpa/Opengate proteasome complex for up to 24 h. PrcB is the proteasomal β-subunit, PreB is unprocessed proteasomal β-subunit. E. Mpa-Proteasome complex was over-expressed for 16 h, followed by over-expression of Pup-GGQ for up to 12 h. Over-expression of the proteasome complex does not result in the degradation of Pup-GGQ. For quantitation of changes in chemical shifts and peak intensities see Figure S4 .