Reconstitution of Bam Complex-Mediated Assembly of a Trimeric Porin into Proteoliposomes.

ABSTRACT: Many integral membrane proteins form oligomeric complexes, but the assembly of these structures is poorly understood. Here, we show that the assembly of OmpC, a trimeric porin that resides in the Escherichia coli outer membrane (OM), can be reconstituted in vitro. Although we observed the insertion of both urea-denatured and in vitro-synthesized OmpC into pure lipid vesicles at physiological pH, the protein assembled only into dead-end dimers. In contrast, in vitro-synthesized OmpC was inserted into proteoliposomes that contained the barrel assembly machinery (Bam) complex, a conserved heterooligomer that catalyzes protein integration into the bacterial OM, and folded into heat-stable trimers by passing through a short-lived dimeric intermediate. I

SUBMITTER: Hussain S 

PROVIDER: S-EPMC8406188 | biostudies-literature | 2021 Aug

REPOSITORIES: biostudies-literature