title: Diffusion of MMPs on the surface of collagen fibrils: the mobile cell surface-collagen substratum interface.
abstract: Remodeling of the extracellular matrix catalyzed by MMPs is central to morphogenetic phenomena during development and wound healing as well as in numerous pathologic conditions such as fibrosis and cancer. We have previously demonstrated that secreted MMP-2 is tethered to the cell surface and activated by MT1-MMP/TIMP-2-dependent mechanism. The resulting cell-surface collagenolytic complex (MT1-MMP)(2)/TIMP-2/MMP-2 can initiate (MT1-MMP) and complete (MMP-2) degradation of an underlying collagen fibril. The following question remained: What is the mechanism of substrate recognition involving the two structures of relatively restricted mobility, the cell surface enzymatic complex and a collagen fibril embedded in the ECM? Here we demonstrate that all the components of the complex are capable of processive movement on a surface of the collagen fibril. The mechanism of MT1-MMP movement is a biased diffusion with the bias component dependent on the proteolysis of its substrate, not adenosine triphosphate (ATP) hydrolysis. It is similar to that of the MMP-1 Brownian ratchet we described earlier. In addition, both MMP-2 and MMP-9 as well as their respective complexes with TIMP-1 and -2 are capable of Brownian diffusion on the surface of native collagen fibrils without noticeable dissociation while the dimerization of MMP-9 renders the enzyme immobile. Most instructive is the finding that the inactivation of the enzymatic activity of MT1-MMP has a detectable negative effect on the cell force developed in miniaturized 3D tissue constructs. We propose that the collagenolytic complex (MT1-MMP)(2)/TIMP-2/MMP-2 represents a Mobile Cell Surface-Collagen Substratum Interface. The biological implications of MT1-MMP acting as a molecular ratchet tethered to the cell surface in complex with MMP-2 suggest a new mechanism for the role of spatially regulated peri-cellular proteolysis in cell-matrix interactions.
authors.affiliation: Division of Dermatology, Department of Medicine, Washington University School of Medicine, Saint Louis, Missouri, United States of America.
authors.forename: Ivan E
authors.initials: IE
authors.lastname: Collier
authors.forename: Wesley
authors.initials: W
authors.lastname: Legant
authors.forename: Barry
authors.initials: B
authors.lastname: Marmer
authors.forename: Olga
authors.initials: O
authors.lastname: Lubman
authors.forename: Saveez
authors.initials: S
authors.lastname: Saffarian
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Wakatsuki
authors.forename: Elliot
authors.initials: E
authors.lastname: Elson
authors.forename: Gregory I
authors.initials: GI
authors.lastname: Goldberg
mesh_terms: D000818:Animals; D003094:Collagen / Q000737:chemistry* / Q000378:metabolism*; D004058:Diffusion; D004220:Disulfides / Q000737:chemistry; D004789:Enzyme Activation; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D005110:Extracellular Space / Q000378:metabolism; D005780:Gelatin / Q000737:chemistry; D006801:Humans; D020782:Matrix Metalloproteinases / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D009068:Movement; D011485:Protein Binding; D059748:Proteolysis; D051381:Rats
chemical_list: D004220:Disulfides; D004800:Enzymes, Immobilized; D005780:Gelatin; D003094:Collagen; D020782:Matrix Metalloproteinases