Identification of surface proteins by shaving of the surface of live cells and LC/MS/MS analysis. Bacteria were grown at mid-exponential phase and harvested by centrifugation. After washing in PBS, they were resuspended in incubation buffer and digested with a protease. Supernatants containing the released peptides were recovered, and an aliquot was re-digested with the same protease to make remaining large polypeptides more amenable to LC/MS/MS analysis. Pellets consisting of the shaved bacteria were plated onto blood-agar plates to test cell viability.
Rodríguez-Ortega et al. BMC Genomics 2008 9:588 doi:10.1186/1471-2164-9-588