Methods of identifying membrane proteins in spirochetes
Abstract
Bacterial membrane proteins serve vital functions such as nutrient acquisition, sensation of the environment, and in gene regulation, secretion, and attachment. Proteins on the cell surface are instrumental in host-pathogen interactions and many serve as immunogens that confer protection as targets for neutralizing antibodies. Integral membrane and lipidated proteins possess hydrophobic domains or lipid anchors that interact with the lipid bilayer of cellular membranes, allowing the investigator to use hydrophobicity as a means for enrichment. The spirochete Borrelia burgdorferi is a Gram negative-like microorganism that produces many integral and lipidated proteins, several of which have proven important during infection and transmission of the bacterium from the tick vector to the mammalian host. Protocols described in this unit for enriching membrane proteins have been extensively used by investigators in the study of B. burgdorferi, but can be easily adapted to identify and characterize membrane-associated and surface-exposed proteins associated with other bacteria.