A Molecularly Complete Planar Bacterial Outer Membrane Platform.
The bacterial outer membrane (OM) is a barrier containing membrane proteins and liposaccharides that fulfill crucial functions for Gram-negative bacteria. With the advent of drug-resistant bacteria, it is necessary to understand the functional role of this membrane and its constituents to enable novel drug designs. Here we report a simple method to form an OM-like supported bilayer (OM-SB), which incorporates native lipids and membrane proteins of gram-negative bacteria from outer membrane vesicles (OMVs). We characterize the formation of OM-SBs using quartz crystal microbalance with dissipation (QCM-D) and fluorescence microscopy. We show that the orientation of proteins in the OM-SB matches the native bacterial membrane, preserving the characteristic asymmetry of these membranes. As a demonstration of the utility of the OM-SB platform, we quantitatively measure antibiotic interactions between OM-SBs and polymyxin B, a cationic peptide used to treat Gram-negative infections. This data enriches understanding of the antibacterial mechanism of polymyxin B, including disruption kinetics and changes in membrane mechanical properties. Combining OM-SBs with microfluidics will enable higher throughput screening of antibiotics. With a broader view, we envision that a molecularly complete membrane-scaffold could be useful for cell-free applications employing engineered membrane proteins in bacterial membranes for myriad technological purposes.
School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY, USA.
Chih-Yun
CY
Hsia
School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY, USA.
Linxiao
L
Chen
School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY, USA.
Rohit R
RR
Singh
School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY, USA.
Matthew P
MP
DeLisa
School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY, USA.
Susan
S
Daniel
D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D007700:Kinetics; D008081:Liposomes; D008856:Microscopy, Fluorescence; D058525:Quartz Crystal Microbalance Techniques
D001425:Bacterial Outer Membrane Proteins; D008081:Liposomes