Discovery of a distinct BAM complex in the Bacteroidetes

Silale, A.; Madej, M.; Mikruta, K.; Frey, A. M.; Hart, A. J.; Basle, A.; Scavenius, C.; Enghild, J. J.; Trost, M.; Hirt, R. P.; van den Berg, B.

doi:10.1101/2025.01.31.636011

bioRxiv · 10.1101/2025.01.31.636011

Discovery of a distinct BAM complex in the Bacteroidetes

Silale, A.·Madej, M.·Mikruta, K.·Frey, A. M.·Hart, A. J.·Basle, A.·Scavenius, C.·Enghild, J. J.·Trost, M.·Hirt, R. P.·van den Berg, B.

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Abstract

The BAM ({beta}-barrel assembly machinery) complex is an evolutionarily conserved, multiprotein machine that catalyses the folding and membrane insertion of newly synthesised {beta}-barrel outer membrane (OM) proteins in Gram-negative bacteria. Based on Proteobacteria, bacterial BAM is also structurally conserved, with an essential BamAD core and up to three auxiliary periplasmic lipoproteins of poorly defined function. Here we show, using structural biology, quantitative proteomics and functional assays, that the BAM complex is radically different within the Bacteroidetes, a large and important phylum widely distributed within the environment and animal microbiomes. Cryogenic electron microscopy (cryo-EM) structures of BAM complexes from the human gut symbiont Bacteroides thetaiotaomicron and the human oral pathogen Porphyromonas gingivalis show similar, seven-component complexes of [~]325 kDa in size with most of the mass in the extracellular space. In addition to canonical BamA and BamD, the complexes contain an integral OM protein named BamF that is essential and intimately associated with BamA, as well as four surface-exposed lipoproteins (SLPs) named BamG-J. Together, BamF-J form a large, extracellular dome that likely serves as an assembly cage for the {beta}-barrel-SLP complexes that are a hallmark of the Bacteroidetes. Our data suggest that BAM functionality in Bacteroidetes is substantially expanded from that in Proteobacteria and underscores the importance of studying other phyla for a more complete understanding of fundamental biological processes.

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Silale, A., Madej, M., Mikruta, K., Frey, A. M., Hart, A. J., Basle, A., Scavenius, C., Enghild, J. J., Trost, M., Hirt, R. P., van den Berg, B.. 2025-02-01. Discovery of a distinct BAM complex in the Bacteroidetes. https://doi.org/10.1101/2025.01.31.636011

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