title: Elastase and tryptase govern TNFα-mediated production of active chemerin by adipocytes.
abstract: Chemerin is a leukocyte chemoattractant and adipokine with important immune and metabolic roles. Chemerin, secreted in an inactive form prochemerin, undergoes C-terminal proteolytic cleavage to generate active chemerin, a ligand for the chemokine-like receptor-1 (CMKLR1). We previously identified that adipocytes secrete and activate chemerin. Following treatment with the obesity-associated inflammatory mediator TNFα, unknown adipocyte mechanisms are altered resulting in an increased ratio of active to total chemerin production. Based on these findings we hypothesized adipocytes produce proteases capable of modifying chemerin and its ability to activate CMKRL1. 3T3-L1 adipocytes expressed mRNA of immunocyte and fibrinolytic proteases known to activate chemerin in vitro. Following treatment with a general protease inhibitor cocktail (PIC), the TNFα-stimulated increase in apparent active chemerin concentration in adipocyte media was amplified 10-fold, as measured by CMKLR1 activation. When the components of the PIC were investigated individually, aprotinin, a serine protease inhibitor, blocked 90% of the TNFα-associated increase in active chemerin. The serine proteases, elastase and tryptase were elevated in adipocyte media following treatment with TNFα and their targeted neutralization recapitulated the aprotinin-mediated effects. In contrast, bestatin, an aminopeptidase inhibitor, further elevated the TNFα-associated increase in active chemerin. Our results support that adipocytes regulate chemerin by serine protease-mediated activation pathways and aminopeptidase deactivation pathways. Following TNFα treatment, increased elastase and tryptase modify the balance between activation and deactivation, elevating active chemerin concentration in adipocyte media and subsequent CMKLR1 activation.
authors.affiliation: Department of Pharmacology, Dalhousie University, Halifax, Nova Scotia, Canada.
authors.forename: Sebastian D
authors.initials: SD
authors.lastname: Parlee
authors.forename: Jenna O
authors.initials: JO
authors.lastname: McNeil
authors.forename: Shanmugam
authors.initials: S
authors.lastname: Muruganandan
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Sinal
authors.forename: Kerry B
authors.initials: KB
authors.lastname: Goralski
mesh_terms: D041721:3T3-L1 Cells; D017667:Adipocytes / Q000187:drug effects / Q000201:enzymology*; D000818:Animals; D018925:Chemokines; D002630:Chemotactic Factors / Q000096:biosynthesis*; D003470:Culture Media / Q000494:pharmacology; D005342:Fibrinolysis / Q000187:drug effects; D036341:Intercellular Signaling Peptides and Proteins / Q000096:biosynthesis*; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D008214:Lymphocytes / Q000187:drug effects / Q000201:enzymology; D051379:Mice; D008810:Mice, Inbred C57BL; D008954:Models, Biological; D010196:Pancreatic Elastase / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology; D059748:Proteolysis / Q000187:drug effects; D019707:Receptors, Chemokine; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D053802:Tryptases / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
chemical_list: C486456:CMKLR1 protein, mouse; D018925:Chemokines; D002630:Chemotactic Factors; D003470:Culture Media; D036341:Intercellular Signaling Peptides and Proteins; D011480:Protease Inhibitors; D019707:Receptors, Chemokine; D043562:Receptors, G-Protein-Coupled; D014409:Tumor Necrosis Factor-alpha; C523686:chemerin protein, mouse; D010196:Pancreatic Elastase; D053802:Tryptases; D007930:Leucine; C012211:ubenimex