Affinity labeling and partial purification of nerve growth factor receptors from rat pheochromocytoma and human melanoma cells.
A cell surface glycoprotein receptor for nerve growth factor (NGF) has been identified by covalent crosslinking to 125I-labeled NGF (125I-NGF). Either ethyldimethylisopropyl-aminocarbodiimide or hydroxysuccinimidyl-p-azidobenzoate causes highly specific crosslinking of 125I-NGF to a similar receptor species on rat pheochromocytoma PC12 cells and on human melanoma A875 cells. The NGF-receptor complex migrates as a broad band in NaDodSO4/polyacrylamide gel electrophoresis with an apparent Mr of approximately equal to 100,000. Because the NaDodSO4-denatured complex apparently contains a single Mr 13,000 NGF chain, the apparent molecular weight of the receptor itself is 87,000. Inhibition of protein glycosylation by tunicamycin generates smaller 125I-NGF-receptor complexes. The mobility of the smallest of these in NaDodSO4 gel electrophoresis corresponds to a Mr of 90,000 for the complex and, hence, 77,000 for the carbohydrate-deficient receptor. A second NGF-receptor complex with a Mr of 225,000 also is obtained from A875 cells but only occasionally from PC12 cells. Tunicamycin treatment decreases the molecular weight of these species by 10,000-15,000. Substantial purification of the Mr 100,000 NGF-receptor complex was achieved by lectin affinity chromatography on columns of wheat germ agglutinin linked to Affi-Gel 15. The specific absorption of NGF-receptor complexes to these columns indicates that the receptor is a glycoprotein that contains N-acetyl-D-glucosamine.
D000310:Adrenal Gland Neoplasms / Q000378:metabolism*; D000345:Affinity Labels / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D006801:Humans; D008545:Melanoma / Q000378:metabolism*; D008970:Molecular Weight; D009414:Nerve Growth Factors / Q000378:metabolism*; D010673:Pheochromocytoma / Q000378:metabolism*; D051381:Rats; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D017475:Receptors, Nerve Growth Factor
D000345:Affinity Labels; D003432:Cross-Linking Reagents; D009414:Nerve Growth Factors; D011956:Receptors, Cell Surface; D017475:Receptors, Nerve Growth Factor