Characterization of a laminin receptor on rat hepatocytes.
The interaction of rat hepatocytes with laminin was studied. The cells were found to adhere to the distal half of the long arm in the laminin molecule (fragment E8), in addition to the previously identified site in the central cross of laminin (fragment P1). Attachment to laminin and to each of the two cell-binding fragments was inhibited by antibodies against the integrin beta 1-subunit of the fibronectin receptor, but not by the cell-binding peptide of fibronectin (Gly-Arg-Gly-Asp-Ser-Cys). By affinity chromatography on laminin-Sepharose in the presence of 2 mM Mn2+, the beta 1-subunit was isolated together with an alpha-subunit with an unreduced Mr of 180,000. This laminin-binding integrin did not bind to Sepharose conjugated with a 105-kDa cell-binding fragment of fibronectin and conversely, the fibronectin receptor of the cells (integrin alpha 5 beta 1) did not bind to the laminin-Sepharose. The 180-kDa protein was identified as the integrin subunit alpha 1 based on its specific reactivity with antibodies raised against a peptide of the N-terminal part of human alpha 1. Integrin alpha 1 beta 1 was found to bind at physiological ionic strength also to Sepharose conjugated with either one of the laminin fragments P1 or E8. Furthermore, integrin alpha 1 beta 1 isolated on one of the fragment columns could be shown to rebind to the other fragment-Sepharose. The results indicate that two structurally distinct domains of laminin may interact with the same type of receptor on hepatocytes.
Forsberg
E
E
Department of Medical and Physiological Chemistry, University of Uppsala, Sweden.
Paulsson
M
M
Timpl
R
R
Johansson
S
S
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D000936:Antigen-Antibody Complex; D002462:Cell Membrane / Q000276:immunology; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D006801:Humans; D007074:Immunoglobulin G; D007700:Kinetics; D007797:Laminin / Q000378:metabolism*; D008099:Liver / Q000276:immunology*; D046911:Macromolecular Substances; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D051381:Rats; D011919:Rats, Inbred Strains; D011971:Receptors, Immunologic / Q000302:isolation & purification / Q000378:metabolism*; D017474:Receptors, Laminin; D012689:Sequence Homology, Nucleic Acid
D000906:Antibodies; D000936:Antigen-Antibody Complex; D007074:Immunoglobulin G; D007797:Laminin; D046911:Macromolecular Substances; D011971:Receptors, Immunologic; D017474:Receptors, Laminin