title: Different beta 1-integrin collagen receptors on rat hepatocytes and cardiac fibroblasts.
abstract: Detergent extracts of primary rat hepatocytes and neonatal cardiac fibroblasts were applied to collagen type I-Sepharose in the presence of 1 mM MnCl2. Elution of bound proteins by 10 mM EDTA yielded one beta 1-integrin heterodimer from hepatocytes with an Mr of 180,000/115,000 under nonreducing conditions. Two beta 1-integrins with Mr's (nonreduced) of 180,000/115,000 and 145,000/115,000 could be isolated from surface-iodinated fibroblasts. A monoclonal antibody, 3A3, directed against the rat homolog of the human integrin VLA-1, precipitated the affinity-purified Mr 180,000/115,000 heterodimer, establishing the relatedness of the Mr 180,000 subunit to the alpha 1-chain of the beta 1-integrin subfamily. Both the alpha 1 beta 1-integrin and the 145,000/beta 1-integrin heterodimers bound specifically to Sepharose beads derivatized with the collagen fragment alpha 1(I) CB3, which lacks RGD sequences. Immunofluorescence staining using the 3A3 monoclonal antibody revealed that the rat alpha 1 beta 1-integrin was present at focal adhesion sites of fibroblasts grown on native collagen type I- but not on fibronectin-coated substrates, although both types of substrates supported the formation of beta 1-integrin containing focal adhesions. Similarly, hepatocytes cultured on substrata coated with collagen type I (but not fibronectin) were stained in a patchy pattern localized to the cell periphery by 3A3 IgG. Furthermore, 3A3 IgG completely inhibited the attachment of hepatocytes to collagen type I, whereas under identical conditions the attachment of fibroblasts to these substrates was inhibited only by approximately 40%. The attachment of both hepatocytes and cardiac fibroblasts to fibronectin was unaffected by the presence of the 3A3 antibody. Collectively these data show that a rat homolog of the human VLA-1 heterodimer both biochemically and functionally fulfills the criteria of a single collagen receptor on rat hepatocytes. In contrast, rat cardiac fibroblasts utilize two different collagen-binding integrins to adhere to collagen, one of which is the rat homolog of the human VLA-1 heterodimer. Furthermore alpha 1(I) CB3 contains cell binding sites for beta 1-integrins.
authors.affiliation: Department of Medical and Physiological Chemistry, Uppsala University, Sweden.
authors.forename: D
authors.initials: D
authors.lastname: Gullberg
authors.forename: D C
authors.initials: DC
authors.lastname: Turner
authors.forename: T K
authors.initials: TK
authors.lastname: Borg
authors.forename: L
authors.initials: L
authors.lastname: Terracio
authors.forename: K
authors.initials: K
authors.lastname: Rubin
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism / Q000502:physiology; D004492:Edetic Acid; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000378:metabolism / Q000648:ultrastructure*; D005455:Fluorescent Antibody Technique; D016023:Integrins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D008099:Liver / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D009206:Myocardium / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D009842:Oligopeptides / Q000032:analysis / Q000378:metabolism; D051381:Rats; D011956:Receptors, Cell Surface / Q000302:isolation & purification / Q000378:metabolism*; D038981:Receptors, Collagen
chemical_list: D016023:Integrins; D009842:Oligopeptides; D011956:Receptors, Cell Surface; D038981:Receptors, Collagen; C047981:arginyl-glycyl-aspartic acid; D003094:Collagen; D004492:Edetic Acid