Matrix metalloproteinase processing of monocyte chemoattractant proteins generates CC chemokine receptor antagonists with anti-inflammatory properties in vivo.
Monocyte chemoattractant protein (MCP)-3 is inactivated upon cleavage by the matrix metalloproteinase (MMP) gelatinase A (MMP-2). We investigated the susceptibility to proteolytic processing of the 4 human MCPs by 8 recombinant MMPs to determine whether MCP-3 is an isolated example or represents a general susceptibility of chemokines to proteolytic inactivation by these important inflammatory proteases. In addition to MMP-2, MCP-3 is efficiently cleaved by membrane type 1 (MT1)-MMP, the cellular activator of MMP-2, and by collagenase-1 and collagenase-3 (MMP-1, MMP-13) and stromelysin-1 (MMP-3). Specificity was shown by absence of cleavage by matrilysin (MMP-7) and the leukocytic MMPs neutrophil collagenase (MMP-8) and gelatinase B (MMP-9). The closely related chemokines MCP-1, MCP-2, and MCP-4 were not cleaved by MMP-2 or MT1-MMP, but were cleaved by MMP-1 and MMP-3 with varying efficiency. MCPs were typically cleaved between residues 4 and 5, but MCP-4 was further processed at Val7-Pro8. Synthetic MCP analogs corresponding to the MMP-cleaved forms bound CC chemokine receptor (CCR)-2 and CCR-3, but lacked chemoattractant activity in pre-B cells transfected with CCR-2 and CCR-3 or in THP-1 monocytic cells, a transformed leukemic cell line. Moreover, the truncated products of MCP-2 and MCP-4, like MCP-3, were potent antagonists of their cognate CC chemokine receptors in transwell cell migration assays in vitro. When they were injected 24 hours after the initiation of carrageenan-induced inflammation in rat paws, their in vivo antagonist activities were revealed by a greater than 66% reduction in inflammatory edema progression after 12 hours. We propose that MMPs have an important role in modulating inflammatory and immune responses by processing chemokines in wound healing and in disease.
McQuibban
G Angus
GA
Department of Biochemistry and Molecular Biology and the Biomedical Research Centre, University of British Columbia, Vancouver, Canada.
Gong
Jiang-Hong
JH
Wong
Julie P
JP
Wallace
John L
JL
Clark-Lewis
Ian
I
Overall
Christopher M
CM
D000595:Amino Acid Sequence; D000818:Animals; D000893:Anti-Inflammatory Agents; D001402:B-Lymphocytes / Q000378:metabolism; D016466:CHO Cells; D002351:Carrageenan; D018932:Chemokine CCL2 / Q000378:metabolism; D054410:Chemokine CCL7; D054412:Chemokine CCL8; D017364:Collagenases / Q000378:metabolism; D006224:Cricetinae; D016207:Cytokines; D004487:Edema / Q000139:chemically induced / Q000188:drug therapy; D004789:Enzyme Activation; D005260:Female; D006801:Humans; D007249:Inflammation / Q000139:chemically induced / Q000188:drug therapy; D008297:Male; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism; D053509:Matrix Metalloproteinase 13; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D019278:Matrix Metalloproteinase 3 / Q000378:metabolism; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases / Q000378:metabolism; D018945:Monocyte Chemoattractant Proteins / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D019707:Receptors, Chemokine / Q000037:antagonists & inhibitors* / Q000235:genetics; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity; D014162:Transfection
D000893:Anti-Inflammatory Agents; C515450:CCL7 protein, human; C515453:CCL8 protein, human; C515452:Ccl7 protein, rat; D018932:Chemokine CCL2; D054410:Chemokine CCL7; D054412:Chemokine CCL8; D016207:Cytokines; D018945:Monocyte Chemoattractant Proteins; D010446:Peptide Fragments; D019707:Receptors, Chemokine; D011994:Recombinant Proteins; D002351:Carrageenan; D017364:Collagenases; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13; D020782:Matrix Metalloproteinases; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases; C507083:Mmp13 protein, rat; D019278:Matrix Metalloproteinase 3; D020778:Matrix Metalloproteinase 2; D020781:Matrix Metalloproteinase 1