Molecular characterization of fibronectin binding proteins in bacteria

While it is becoming widely recognized that may different bacterial species interact with the extracellular matrix, little information has been generated relating to the molecular details of these interactions. This review will present methods for the analysis of bacterial binding to the extracellular matrix protein fibronectin. Binding to Streptococcus pyogenes and Escherichia coli will be emphasized to provide specidic examples of how these methods can be applied to the analysis of binding in Gram-positive and Gram-negative bacterial species. Assays for the quantitative measurement of binding will be presented as well as criteria for evaluating binding specificity, affinity and the limitations to these methods. Also, methods will be outlined for cloning and mutagenesis of putative binding proteins to provide a strategy for the molecular analysis of fibronectin binding. These methods should have wide application for the study of binding in many different microbial species and for the analysis of the relevance of fibronectin binding in the context of bacteria-host interplay.