Updated Methods for Elucidating Bacterial Surfaceomes.

The bacterial surfaceome, representing the full complement of surface-expressed proteins, is integral to interactions between bacteria, their hosts, and the environment, influencing processes such as adhesion, invasion, immune evasion, and biofilm formation. These proteins, due to their accessibility and vital roles in bacterial physiology, are prime targets for therapeutic intervention. This protocol describes a detailed methodology for mapping bacterial surfaceomes, including enzymatic surface shaving, cell surface biotinylation, solid-phase extraction, and liquid chromatography-tandem mass spectrometry.