Forster Resonance Energy Transfer Measurements in Living Bacteria for Interaction Studies of BamA with BamD and Inhibitor Identification.

ABSTRACT: The β-barrel assembly machinery (BAM) is a multimeric protein complex responsible for the folding of outer membrane proteins in gram-negative bacteria. It is essential for cell survival and outer membrane integrity. Therefore, it is of impact in the context of antibiotic resistance and can serve as a target for the development of new antibiotics. The interaction between two of its subunits, BamA and BamD, is essential for its function. Here, a FRET-based assay to quantify the affinity between these two proteins in living bacterial cells is presented. The method was applied to identify two interaction hotspots at the binding interface. BamD^Y184 was identified to significantly contribute to the binding between both proteins through hydrophobic interactions and hydrogen bonding. Ad

SUBMITTER: Schreiber S 

PROVIDER: S-EPMC11592675 | biostudies-literature | 2024 Nov

REPOSITORIES: biostudies-literature