Mapping the Binding Sites of MMPs on Types II and III Collagens Using Triple-Helical Peptide Toolkits

Libraries of triple-helical collagen-like peptides (Collagen Toolkits) have helped to define collagens II and III binding specificities of numerous collagen-binding proteins. Here I describe a simple solid-phase binding assay utilizing a biotin–streptavidin system to screen the Collagen Toolkits for binding of two distinct matrix metalloproteinases (MMPs) implicated in cancer: the collagenolytic MMP1 (collagenase 1) and the non-collagenolytic MMP3 (stromelysin 1). The screening revealed markedly disparate binding footprints of these MMPs on collagens II and III, in line with their distinct biological activities. Analogous screening of other potentially collagen-binding proteases may shed light on their inherent tissue retention capabilities and their pro- or anti-metastatic potential.