Journal article
The WD40 Protein BamB Mediates Coupling of BAM Complexes into Assembly Precincts in the Bacterial Outer Membrane
SD Gunasinghe, T Shiota, CJ Stubenrauch, KE Schulze, CT Webb, AJ Fulcher, RA Dunstan, ID Hay, T Naderer, DR Whelan, TDM Bell, KD Elgass, RA Strugnell, T Lithgow
Cell Reports | CELL PRESS | Published : 2018
Open access
Abstract
The β-barrel assembly machinery (BAM) complex is essential for localization of surface proteins on bacterial cells, but the mechanism by which it functions is unclear. We developed a direct stochastic optical reconstruction microscopy (dSTORM) methodology to view the BAM complex in situ. Single-cell analysis showed that discrete membrane precincts housing several BAM complexes are distributed across the E. coli surface, with a nearest neighbor distance of ∼200 nm. The auxiliary lipoprotein subunit BamB was crucial for this spatial distribution, and in situ crosslinking shows that BamB makes intimate contacts with BamA and BamB in neighboring BAM complexes within the precinct. The BAM complex..
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Grants
Awarded by Australian Research Council
Funding Acknowledgements
The authors thank Bruker for access to the Vutara SR350 super-resolution system and, particularly, Carl Ebeling (Bruker) for assistance with the system. We thank Franz-Ulrich Hartl for expert advice on quantative proteomics, David Steer for expertise in protein mass spectrometry (Monash Biomedical Proteomics Facility), and Jonathan Wilksch for comments on the manuscript. This work was supported by the Multi-modal Australian Sciences Imaging and Visualisation Environment (MASSIVE; www. massive. org. au) and funded through an Australian Research Council (ARC) Laureate fellowship award (FL30100038), an ARC LIEF grant (LE150100110), and an NHMRC program grant (1092262). T. L. is an ARC Australian Laureate fellow, I. D. H. is an ARC Laureate postdoctoral fellow, and S. D. G. is the recipient of an ARC Laureate postgraduate research scholarship.