Characterization of the human melanoma nerve growth factor receptor.
Monoclonal antibodies to the human nerve growth factor receptor have been used to biochemically characterize the receptor in the human melanoma cell line A875. Labeling of A875 cell proteins by culture with [35S]cysteine or labeling of cell surface proteins with 125I followed by immunoprecipitation with anti-nerve growth factor receptor antibody reveals a receptor protein with an apparent Mr of 70,000-75,000 and an isoelectric point of 4.9-5.2. Incorporation of [3H]glucosamine into this species indicates it is a glycoprotein. The receptor becomes phosphorylated on serine residues in intact cells and in isolated membranes incubated with [gamma-32P]ATP. The receptor appears to exist, at least partially, in the form of a disulfide-linked oligomer (probably a dimer) of Mr = 75,000 subunits. Kinetic [35S]cysteine labeling studies reveal an Mr = 59,000 core protein which is glycosylated via N-linked and probably also O-linked sugar moieties to produce the mature (Mr = 70,000-75,000) receptor.
Grob
P M
PM
Ross
A H
AH
Koprowski
H
H
Bothwell
M
M
D000911:Antibodies, Monoclonal; D002460:Cell Line; D003545:Cysteine / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005944:Glucosamine / Q000378:metabolism; D006801:Humans; D008545:Melanoma / Q000032:analysis*; D008623:Mercaptoethanol / Q000494:pharmacology; D008970:Molecular Weight; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D011956:Receptors, Cell Surface / Q000032:analysis*; D017475:Receptors, Nerve Growth Factor
D000911:Antibodies, Monoclonal; D011956:Receptors, Cell Surface; D017475:Receptors, Nerve Growth Factor; D008623:Mercaptoethanol; D003545:Cysteine; D005944:Glucosamine