title: Single-molecule tracking of collagenase on native type I collagen fibrils reveals degradation mechanism.
abstract: BACKGROUND
  Collagen, the most abundant human protein, is the principal component of the extracellular matrix and plays important roles in maintaining tissue and organ integrity. Highly resistant to proteolysis, fibrillar collagen is degraded by specific matrix metalloproteases (MMPs). Degradation of fibrillar collagen underlies processes including tissue remodeling, wound healing, and cancer metastasis. However, the mechanism of native collagen fibril degradation remains poorly understood.
  
  
  RESULTS
  Here we present the results of high-resolution tracking of individual MMPs degrading type I collagen fibrils. MMP1 exhibits cleavage-dependent biased and hindered diffusion but spends 90% ± 3% of the time in one of at least two distinct pause states. One class of exponentially distributed pauses (class I pauses) occurs randomly along the fibril, whereas a second class of pauses (class II pauses) exhibits multistep escape kinetics and occurs periodically at intervals of 1.3 ± 0.2 μm and 1.5 ± 0.2 μm along the fibril. After these class II pauses, MMP1 moved faster and farther in one direction along the fibril, indicative of biased motion associated with cleavage. Simulations indicate that 5% ± 2% of the class II pauses result in the initiation of processive collagen degradation, which continues for bursts of 15 ± 4 consecutive cleavage events.
  
  
  CONCLUSIONS
  These findings provide a mechanistic paradigm for type I collagen degradation by MMP1 and establish a general approach to investigate MMP-fibrillar collagen interactions. More generally, this work demonstrates the fundamental role of enzyme-substrate interactions including binding and motion in determining the activity of an enzyme on an extended substrate.
authors.affiliation: Laboratory of Molecular Biophysics, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Susanta K
authors.initials: SK
authors.lastname: Sarkar
authors.forename: Barry
authors.initials: B
authors.lastname: Marmer
authors.forename: Gregory
authors.initials: G
authors.lastname: Goldberg
authors.forename: Keir C
authors.initials: KC
authors.lastname: Neuman
mesh_terms: D000818:Animals; D024042:Collagen Type I / Q000378:metabolism*; D004058:Diffusion; D005453:Fluorescence; D007700:Kinetics; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism*; D008954:Models, Biological; D056004:Molecular Dynamics Simulation; D059748:Proteolysis; D051381:Rats; D013623:Tail / Q000166:cytology; D013696:Temperature; D013997:Time Factors
chemical_list: D024042:Collagen Type I; D020781:Matrix Metalloproteinase 1