Subunit structure of a laminin-binding integrin and localization of its binding site on laminin.
A laminin receptor was isolated from human MG-63 osteosarcoma cells by affinity chromatography on human laminin. The isolated receptor was defined as the alpha 3 beta 1 integrin by immunoprecipitation with subunit-specific antibodies. A previously unclassified laminin-binding integrin from rat cells was shown also to contain the alpha 3 subunit. Both receptors bound to human and mouse laminin in a radioreceptor assay. They also both bound to some extent to fibronectin in this assay, but only the MG-63 cell receptor showed binding to type IV collagen. The binding of the radiolabeled receptor to insoluble laminin was inhibited by unlabeled receptor, by soluble laminin, and by chymotryptic fragments of laminin that have previously been shown to contain neurite-promoting and cell attachment-promoting activities. Moreover, the receptor binding was also inhibited by monoclonal antibodies capable of inhibiting the neurite-promoting activity of laminin and known to bind to laminin near the junction of the long arm and its terminal globule. One of these antibodies was reactive with fusion proteins expressed from laminin cDNA clones. The immunoreactive clones corresponded to the COOH-terminal end of the B1 subunit. These results identify the integrin-type laminin receptor isolated from the osteosarcoma cells as the alpha 3 beta 1 integrin and localize its binding site in close proximity of the B1 subunit COOH terminus.
Gehlsen
K R
KR
Cancer Research Center, La Jolla Cancer Research Foundation, California 92037.
Dickerson
K
K
Argraves
W S
WS
Engvall
E
E
Ruoslahti
E
E
D001665:Binding Sites; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D016023:Integrins / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D007797:Laminin / Q000378:metabolism*; D046911:Macromolecular Substances; D008970:Molecular Weight; D012516:Osteosarcoma; D011869:Radioligand Assay
D016023:Integrins; D007797:Laminin; D046911:Macromolecular Substances