Modeling intermediates of BamA folding an outer membrane protein.

ABSTRACT

BamA, the core component of the ß-barrel assembly machinery complex, is an integral outer-membrane protein (OMP) in Gram-negative bacteria that catalyzes the folding and insertion of OMPs. A key feature of BamA relevant to its function is a lateral gate between its first and last ß-strands. Opening of this lateral gate is one of the first steps in the asymmetric-hybrid-barrel model of BamA function. In this study, multiple hybrid-barrel folding intermediates of BamA and a substrate OMP, EspP, were constructed and simulated to better understand the model's physical consequences. The hybrid-barrel intermediates consisted of the BamA ß-barrel and its POTRA5 domain and either one, two, three, four, five, or six ß-hairpins of EspP. The simulation results support an asymmetric-hybrid-barrel model in which the BamA N-terminal ß-strand forms stronger interactions with the substrate OMP than the C-terminal ß-strand. A consistent "B"-shaped conformation of the final folding intermediate was observed, and the shape of the substrate ß-barrel within the hybrid matched the shape of the fully folded substrate. Upon further investigation, inward-facing glycines were found at sharp bends within the hybrid and fully folded ß-barrels. Together, the data suggest an influence of sequence on shape of the substrate barrel throughout the OMP folding process and of the fully folded OMP.