Proteomic identification of in vivo substrates for matrix metalloproteinases 2 and 9 reveals a mechanism for resolution of inflammation

Authors

Kendra J Greenlee
David B Corry
David A Engler
Risë K Matsunami
Philippe Tessier
Richard G Cook
Zena Werb
Farrah Kheradmand

Publication Date

11-15-2006

Journal

Journal of Immunology

Abstract

Clearance of allergic inflammatory cells from the lung through matrix metalloproteinases (MMPs) is necessary to prevent lethal asphyxiation, but mechanistic insight into this essential homeostatic process is lacking. In this study, we have used a proteomics approach to determine how MMPs promote egression of lung inflammatory cells through the airway. MMP2- and MMP9-dependent cleavage of individual Th2 chemokines modulated their chemotactic activity; however, the net effect of complementing bronchoalveolar lavage fluid of allergen-challenged MMP2(-/-)/MMP9(-/-) mice with active MMP2 and MMP9 was to markedly enhance its overall chemotactic activity. In the bronchoalveolar fluid of MMP2(-/-)/MMP9(-/-) allergic mice, we identified several chemotactic molecules that possessed putative MMP2 and MMP9 cleavage sites and were present as higher molecular mass species. In vitro cleavage assays and mass spectroscopy confirmed that three of the identified proteins, Ym1, S100A8, and S100A9, were substrates of MMP2, MMP9, or both. Function-blocking Abs to S100 proteins significantly altered allergic inflammatory cell migration into the alveolar space. Thus, an important effect of MMPs is to differentially modify chemotactic bioactivity through proteolytic processing of proteins present in the airway. These findings provide a molecular mechanism to explain the enhanced clearance of lung inflammatory cells through the airway and reveal a novel approach to target new therapies for asthma.

Keywords

Amino Acid Sequence, Animals, Bronchoalveolar Lavage Fluid, Calgranulin B, Cell Migration Inhibition, Chemotaxis, Leukocyte, Dose-Response Relationship, Immunologic, Hydrolysis, Inflammation Mediators, Lectins, Lung, Matrix Metalloproteinase 2, Matrix Metalloproteinase 9, Mice, Mice, Inbred C57BL, Mice, Knockout, Molecular Sequence Data, Proteome, Respiratory Hypersensitivity, S100 Proteins, Substrate Specificity, Th2 Cells, beta-N-Acetylhexosaminidases