Poster Presentation The 43rd Lorne Conference on Protein Structure and Function 2018

Beta-barrel assembly machinery (BAM) in action; a spatiotemporal characterisation using Single Molecule Localization Microscopy (SMLM). (#133)

Sachith D Gunasinghe 1 , Trevor Lithgow 1 , Kirstin Elgass 2 , Toby Bell 3
  1. Department of Microbiology, Monash University, Clayton, VIC, Australia
  2. Hudson Institute of Medical Research, Clayton, VIC, Australia
  3. School of Chemistry, Monash University, Clayton, VIC, Australia

The Beta-barrel assembly machinery (BAM) is essential for localization and function of the vast majority of cell surface proteins on bacterial cells. We developed a direct stochastic optical reconstruction microscopy (dSTORM) methodology and used video-rate, single-molecule localization microscopy to view the BAM complex in situ. Further, we believe the auxiliary lipoprotein subunit BamB is crucial for this spatial distribution. Statistical analysis showed that the BAM complex clusters grow when outer membrane protein synthesis is maximal, visual proof that the precincts are active in protein assembly. This nanoscale interrogation of the BAM complex in situ suggests a new model whereby bacterial outer membranes contain highly organized assembly precincts to drive integral protein assembly.