Control of promatrilysin (MMP7) activation and substrate-specific activity by sulfated glycosaminoglycans.
Matrix metalloproteinases are maintained in an inactive state by a bond between the thiol of a conserved cysteine in the prodomain and a zinc atom in the catalytic domain. Once this bond is disrupted, MMPs become active proteinases and can act on a variety of extracellular protein substrates. In vivo, matrilysin (MMP7) activates pro-alpha-defensins (procryptdins), but in vitro, processing of these peptides is slow, with about 50% conversion in 8-12 h. Similarly, autolytic activation of promatrilysin in vitro can take up to 12-24 h for 50% conversion. These inefficient reactions suggest that natural cofactors enhance the activation and activity of matrilysin. We determined that highly sulfated glycosaminoglycans (GAG), such as heparin, chondroitin-4,6-sulfate (CS-E), and dermatan sulfate, markedly enhanced (>50-fold) the intermolecular autolytic activation of promatrilysin and the activity of fully active matrilysin to cleave specific physiologic substrates. In contrast, heparan sulfate and less sulfated forms of chondroitin sulfate did not augment matrilysin activation or activity. Chondroitin-2,6-sulfate (CS-D) also did not enhance matrilysin activity, suggesting that the presentation of sulfates is more important than the overall degree of sulfation. Surface plasmon resonance demonstrated that promatrilysin bound heparin (K(D), 400 nm) and CS-E (K(D), 630 nm). Active matrilysin bound heparin (K(D), 150 nm) but less so to CS-E (K(D), 60 microm). Neither form bound heparan sulfate. These observations demonstrate that sulfated GAGs regulate matrilysin activation and its activity against specific substrates.
Ra
Hyun-Jeong
HJ
Center for Lung Biology, University of Washington, Seattle, Washington 98109.
Harju-Baker
Susanna
S
Center for Lung Biology, University of Washington, Seattle, Washington 98109.
Zhang
Fuming
F
Department of Chemistry and Chemical Biology and Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, New York 12180.
Linhardt
Robert J
RJ
Department of Chemistry and Chemical Biology and Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, New York 12180.
Wilson
Carole L
CL
Department of Pathology, University of Washington, Seattle, Washington 98109.
Parks
William C
WC
Center for Lung Biology, University of Washington, Seattle, Washington 98109. Electronic address: parksw@u.washington.edu.
D000818:Animals; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000235:genetics / Q000378:metabolism*; D006025:Glycosaminoglycans / Q000737:chemistry / Q000378:metabolism; D006493:Heparin / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism*; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D013431:Sulfates / Q000737:chemistry / Q000378:metabolism*; D020349:Surface Plasmon Resonance
D004792:Enzyme Precursors; D006025:Glycosaminoglycans; D011498:Protein Precursors; D011994:Recombinant Proteins; D013431:Sulfates; C059569:cryptdin; D006493:Heparin; D008666:Metalloendopeptidases; C099583:promatrilysin