Methods for studying mucin-microbe interactions

Mucin glycoproteins are the major structural components of mucus, imparting its gel consistency. Not only do mucins form a protective barrier that traps debris and keeps pathogenic bacteria at a safe distance from host cells, but they display structurally complex glycans that feed beneficial microbes and directly interact with microbial pathogens to regulate gene expression and virulence. This chapter provides a conceptual framework for mucin-microbe interaction studies and integrated protocols spanning the purification of biologically relevant mucins, isolation of complex glycan pools for mechanistic studies, and key experimental approaches for microbiological investigations of mucin function. Together, we hope these protocols will enable the exploration of yet-undiscovered roles of glycans in bacterial physiology and across domains of life.