Molecular mechanism involved in matrix dependent upregulation of matrix metalloproteinases in monocyte/macrophage.
Production of macrophage specific matrix metalloproteinases (MMPs) by monocyte/macrophage (mo/mphi) maintained in vitro on matrix protein substrata has been examined to study the mechanism of matrix protein dependent upregulation of macrophage specific activity. Using specific blocking reagents we have found that interaction of peripheral blood mononuclear cells (PBMC) with extracellular matrix components is crucial for its differentiation to macrophages. Multiwell zymography has shown that production of MMPs was significantly inhibited in cells maintained on fibronectin (FN) pretreated with antibodies to alpha(5), beta(1) integrins and synthetic peptide RGDS. Further, quantification by ELISA showed a significant inhibition in MMP production in cells pretreated with these blocking reagents. Genistein, a non-specific inhibitor of tyrosine kinases, significantly reduced production of MMPs in cells maintained on FN and collagen type IV (COL IV). Immunoblotting analysis has shown that tyrosine phosphorylation occurs in 30 min and two proteins of approximately 115 and approximately 72 kDa are being phosphorylated upon PBMC-FN interaction. These results indicate that integrin mediated downstream signalling involving tyrosine phosphorylation is required for mediating intracellular events associated with differentiation of monocytes to macrophages.
Jacob
Shiney Susan
SS
Department of Biochemistry University of Kerala, Kariavotton, Trivandrum, Kerala 695 581, India.
Sudhakaran
P R
PR
D002454:Cell Differentiation; D002478:Cells, Cultured; D003094:Collagen; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008264:Macrophages / Q000201:enzymology*; D061965:Matrix Metalloproteinase Inhibitors; D020782:Matrix Metalloproteinases / Q000302:isolation & purification / Q000378:metabolism*; D009000:Monocytes / Q000166:cytology / Q000201:enzymology*; D009842:Oligopeptides / Q000494:pharmacology; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism
D061965:Matrix Metalloproteinase Inhibitors; D009842:Oligopeptides; D019000:Phosphotyrosine; D003094:Collagen; C046617:arginyl-glycyl-aspartyl-serine; D020782:Matrix Metalloproteinases