Adhesion of cultured human kidney mesangial cells to native entactin: role of integrin receptors.
Entactin is an extracellular matrix glycoprotein which binds to laminin and is found in most renal basement membranes and in the glomerular mesangial matrix. In the present study, we have characterized specific integrin receptors on cultured human mesangial cells (CHMC) responsible for adhesion to native entactin. The integrin receptors alpha 2 beta 1, alpha 3 beta 1, alpha 5 beta 1, alpha v beta 3, alpha v beta 5, and alpha 6 complexed with either beta 1 or beta 4 could be immune precipitated from detergent extracts of metabolically labeled CHMC. Adhesion assays with inhibitory anti integrin monoclonal antibodies (mab) demonstrated that CHMC use both alpha v beta 3 and a beta 1-containing integrin to bind surfaces coated with native entactin. Optimal binding of CHMC to native entactin required the participation of cations. Using wild type and mutant recombinant entactin fragments, the binding site for the alpha v beta 3 receptor was localized to the RGD sequence on the rod or E domain of entactin. CHMC adhesion to mutant full length recombinant entactin ligands lacking the E domain RGD sequence confirmed the presence of ligand binding site(s) for beta 1 integrin receptor(s). Differences in CHMC binding characteristics to recombinant and full length entactin compared to native bovine basement membrane entactin were observed. This suggests that tertiary molecular structure may contribute to entactin ligand binding properties. Primary amino acid residue sequences and tertiary structure of entactin may play roles in forming functional cell attachment sites in native basement membrane entactin.
Department of Pediatrics, University of Minnesota, Minneapolis 55455, USA.
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Yi
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Wayner
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Kim
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Fish
D000818:Animals; D000911:Antibodies, Monoclonal; D002118:Calcium / Q000378:metabolism; D002412:Cations / Q000378:metabolism; D002417:Cattle; D002448:Cell Adhesion / Q000502:physiology; D002478:Cells, Cultured; D016326:Extracellular Matrix Proteins / Q000378:metabolism / Q000494:pharmacology; D005333:Fetus / Q000166:cytology; D005920:Glomerular Mesangium / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D016023:Integrins / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D008274:Magnesium / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism* / Q000494:pharmacology*; D010446:Peptide Fragments / Q000378:metabolism; D011233:Precipitin Tests
D000911:Antibodies, Monoclonal; D002412:Cations; D016326:Extracellular Matrix Proteins; D016023:Integrins; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; C029967:nidogen; D008345:Manganese; D008274:Magnesium; D002118:Calcium