Monoclonal antibodies to distinctive epitopes on the alpha and beta subunits of the fibronectin receptor.
Monoclonal antibodies (MAbs) have been developed that can recognize epitopes that are unique to either the alpha or beta subunit of the fibronectin receptor (FnR). MAbs 11B4 and 7A8 immunoblot the alpha subunit of FnR either in purified form from Chinese hamster ovary (CHO) cells or in nonionic detergent extracts of cells of human and rodent origin electrophoresed under reducing or nonreducing conditions. The MAbs seem to be more reactive to the subunit when it has been electrophoresed under reducing conditions, suggesting that the epitope may be partially masked by the conformation conferred by disulfide bonding. A second set of MAbs, 7E2 and 7F9, is directed to an epitope on the beta subunit that is conformationally dependent upon disulfide bonding, as reduction of the subunit leads to loss of reactivity with both MAbs. Further, 7E2/7F9 immunoblots of nonionic detergent extracts of CHO cells, run under nonreducing conditions, reveal the presence of a third band (90-kDa), immunologically related to the beta subunit, which is not surface-labeled with 125I in intact cells and which does not copurify with the alpha and beta subunits isolated by immunoaffinity purification of FnR using the MAb PB1. The 90-kDa component is not found associated with a plasma membrane fraction prepared by crude cell fractionation, but is abundant in a low-speed pellet containing nuclei and intracellular membranes. This finding suggests that the 90-kDa component is a precursor to the beta subunit. Finally, the epitope of 7E2/7F9 is unique to CHO cells, as cross-reactivity to other cell types cannot be demonstrated by either immunoblotting or immunoprecipitation.
Brown
P J
PJ
Department of Pharmacology, University of Texas Medical School, Houston 77025.
Juliano
R L
RL
D000818:Animals; D000911:Antibodies, Monoclonal; D002448:Cell Adhesion; D002460:Cell Line; D003429:Cross Reactions; D000939:Epitopes / Q000276:immunology*; D007163:Immunosorbent Techniques; D046911:Macromolecular Substances; D008970:Molecular Weight; D017469:Receptors, Fibronectin; D011971:Receptors, Immunologic / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D046911:Macromolecular Substances; D017469:Receptors, Fibronectin; D011971:Receptors, Immunologic