OmpA Specifically Modulates the Activity of Enzymes that Reside in the Crowded Bacterial Outer Membrane.

The outer membrane (OM) of lipopolysaccharide (LPS) containing-diderm bacteria is crowded with outer membrane proteins (OMPs) that reside in a membrane that is relatively rich in protein and poor in lipid. As a consequence, extensive interactions between OMPs occur. Yet, how these interactions affect OMP function remains unexplored. Here, we examine the effect of OmpA on the activity of three different OMP enzymes, OmpLA (a phospholipase), PagP (a palmitoyltransferase) and OmpT (a protease). We show that OmpA-OmpT interactions enhance the activity of OmpT, and that this catalytic enhancement is mediated via their extracellular loops, an effect that is not observed with other common OMPs, including OmpF and OmpX. In contrast, OmpA specifically reduces the activity of PagP, while OmpLA activity shows no significant change. Possible interactions between the abundant E. coli OMPs (OmpA, OmpF/C, OmpT, OmpX, MipA) and all other E. coli OMPs were screened via Alphafold predictions, with the results suggesting that smaller OMPs are generally more promiscuous interactors, and identifying new interactions that may plausibly form in the OMP-rich islands in the OM. Together, the results identify a previously underappreciated role for specific OMP-OMP interactions in modulating protein function in the OM, and highlight how evolution may have exploited the high local concentrations of abundant OMPs in the OM to tune enzyme activity.