title: Biochemical analysis of matrix metalloproteinase activation of chemokines CCL15 and CCL23 and increased glycosaminoglycan binding of CCL16.
abstract: Leukocyte migration and activation is orchestrated by chemokines, the cleavage of which modulates their activity and glycosaminoglycan binding and thus their roles in inflammation and immunity. Early research identified proteolysis as a means of both activating or inactivating CXC chemokines and inactivating CC chemokines. Recent evidence has shown activating cleavages of the monocyte chemoattractants CCL15 and CCL23 by incubation with synovial fluid, although the responsible proteases could not be identified. Herein we show that CCL15 is processed in human synovial fluid by matrix metalloproteinases (MMPs) and serine proteases. Furthermore, a family-wide investigation of MMP processing of all 14 monocyte-directed CC chemokines revealed that each is precisely cleaved by one or more MMPs. By MALDI-TOF-MS, 149 cleavage sites were sequenced including the first reported instance of CCL1, CCL16, and CCL17 proteolysis. Full-length CCL15-(1-92) and CCL23-(1-99) were cleaved within their unique 31 and 32-amino acid residue extended amino termini, respectively. Unlike other CCL chemokines that lose activity and become receptor antagonists upon MMP cleavage, the prominent MMP-processed products CCL15-(25-92, 28-92) and CCL23-(26-99) are stronger agonists in calcium flux and Transwell CC receptor transfectant and monocytic THP-1 migration assays. MMP processing of CCL16-(1-97) in its extended carboxyl terminus yields two products, CCL16-(8-77) and CCL16-(8-85), with both showing unexpected enhanced glycosaminoglycan binding. Hence, our study reveals for the first time that MMPs activate the long amino-terminal chemokines CCL15 and CCL23 to potent forms that have potential to increase monocyte recruitment during inflammation.
authors.affiliation: Centre for Blood Research, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Amanda E
authors.initials: AE
authors.lastname: Starr
authors.forename: Antoine
authors.initials: A
authors.lastname: Dufour
authors.forename: Josefine
authors.initials: J
authors.lastname: Maier
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Overall
mesh_terms: D000595:Amino Acid Sequence; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D019742:Chemokines, CC / Q000737:chemistry / Q000378:metabolism*; D002633:Chemotaxis; D004789:Enzyme Activation; D006025:Glycosaminoglycans / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008024:Ligands; D019402:Macrophage Inflammatory Proteins / Q000737:chemistry / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D008969:Molecular Sequence Data; D009000:Monocytes / Q000378:metabolism; D011485:Protein Binding; D013379:Substrate Specificity; D013582:Synovial Fluid / Q000166:cytology / Q000378:metabolism
chemical_list: C108814:CCL15 protein, human; C481939:CCL16 protein, human; C105504:CCL23 protein, human; D019742:Chemokines, CC; D006025:Glycosaminoglycans; D008024:Ligands; D019402:Macrophage Inflammatory Proteins; D006493:Heparin; D020782:Matrix Metalloproteinases; D002118:Calcium