Hepatocyte adhesion to collagen. Isolation of membrane glycoproteins involved in adhesion to collagen.
Adhesion of hepatocytes to collagenous substrates and their spreading have been shown to involve a specific recognition event, possibly mediated by membrane proteins with affinity for collagen. In the present communication, we describe the isolation of membrane components that are involved in the adhesion of rat hepatocytes to collagen. These components could be solubilized from liver microsomal membranes by treatment with detergents or papain--but not by treatment with EDTA, urea or high salt. The purification of detergent-solubilized components was monitored by an assay determining the ability of membrane components to neutralize antibody-mediated inhibition of hepatocyte adhesion to collagen. By affinity chromatography on lentil lectin-Sepharose it was found that the neutralizing activity resided within the glycoprotein fraction. These glycoproteins were purified further by affinity-chromatography on collagen type I linked to Sepharose. Antibodies raised against the glycoproteins with affinity for immobilized collagen, effectively inhibited hepatocyte adhesion to collagen. The bulk of the neutralizing activity migrated with an apparent molecular weight of 120 000-140 000 in preparative SDS-PAGE.
D000818:Animals; D000954:Antigens, Surface / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules; D002846:Chromatography, Affinity; D003094:Collagen / Q000378:metabolism*; D006023:Glycoproteins / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D007106:Immune Sera; D008099:Liver / Q000032:analysis / Q000166:cytology* / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D008970:Molecular Weight; D051381:Rats; D011919:Rats, Inbred Strains
D000954:Antigens, Surface; D015815:Cell Adhesion Molecules; D006023:Glycoproteins; D007106:Immune Sera; D008565:Membrane Proteins; D003094:Collagen