Abstract Correct folding of outer membrane proteins (OMPs) by the β-barrel assembly machinery (BAM) is essential for maintaining the outer membrane (OM) barrier function of diderm bacteria. When OMP biogenesis is perturbed, the β-barrel assembly enhancing protease A (BepA) binds to BAM to mediate quality control, but how BepA interacts with BAM and degrades substrate OMPs remains unclear. Here, cryoEM structures of BAM-bound BepA reveals that BepA induces large conformational changes in the BAM complex enabling the enzyme to poise its active site within the periplasmic ring of BAM, beneath the BamA barrel. The lid of BepA is dynamic, embedding two of its water-soluble helices deep into the membrane bilayer when BAM-bound, which readies BepA for proteolysis of misfolding OMPs. Movement of BepA’s plug is triggered by OMP binding rather than interaction with BAM, activating the enzyme for cleavage. We reveal BepA preferentially recognises Aromatic-X-Aromatic (Ar-X-Ar) motifs which are enriched in OMP sequences. The results reveal a mechanism for proteolytic degradation by BepA in OMP quality control which requires interaction with BAM, the membrane, and its OMP substrates.
Fenn et al. (Thu,) studied this question.