Membrane glycoproteins involved in hepatocyte adhesion to collagen type I.
Liver membrane glycoproteins with affinity for immobilized collagen type I were subjected to preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by electroelution of the separated proteins. Electroeluted glycoproteins with ability to neutralize the inhibitory effect of anti-CollCAM antibodies on hepatocyte adhesion to collagen were collected from several consecutive runs and used to raise a high titer antiserum, denoted anti-CollCAM II. IgG from this antiserum inhibited the attachment of hepatocytes to dishes coated with collagen type I, but not to fibronectin- or collagen type IV-coated dishes. When the antibodies were immobilized to Sepharose CL-4B they bound three sets of glycoproteins with apparent Mr's of 105,000, 115,000, and 130,000 as analyzed by SDS-PAGE under nonreducing (NR) conditions. Upon reduction (R) the glycoproteins migrated with apparent Mr's of 115,000, 130,000, and 160,000, respectively. The Mr 105,000-115,000 (NR) glycoproteins effectively neutralized the inhibitory effect exerted by both anti-CollCAM and anti-CollCAM II antibodies, on hepatocyte spreading and attachment to collagen type I substrates. Peptide mapping suggested the Mr 160,000 (R) species to be different from the Mr 115,000 (R).
Gullberg
D
D
Department of Medical and Physiological Chemistry, University of Uppsala, Sweden.
Terracio
L
L
Rubin
K
K
D000818:Animals; D002448:Cell Adhesion; D003094:Collagen / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005353:Fibronectins / Q000378:metabolism; D007118:Immunoassay; D007163:Immunosorbent Techniques; D008099:Liver / Q000166:cytology*; D008562:Membrane Glycoproteins / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D008970:Molecular Weight; D010449:Peptide Mapping
D005353:Fibronectins; D008562:Membrane Glycoproteins; D003094:Collagen