Wheat germ agglutinin, concanavalin A, and lens culinalis agglutinin block the inhibitory effect of nerve growth factor on cell-free phosphorylation of Nsp100 in PC12h cells.
It has been shown that in PC12 and its subclone PC12h treatment of the cells with nerve growth factor (NGF) induces a selective decrease in the incorporation of radioactive phosphate into a 100,000-dalton protein, designated in an earlier study as Nsp100, in the subsequent phosphorylation of soluble extracts from cells with (gamma-32P)ATP. In the present study, we show that plant lectins, wheat germ agglutinin (WGA), concanavalin A (Con A), and lens culinaris agglutinin (LCA), inhibit the action of NGF on Nsp100 phosphorylation in PC12h cells. Treatment of the cells with WGA, which binds to N-acetylglucosamine and sialic acid residues on glycoproteins, strongly blocked the inhibitory action of NGF on the protein phosphorylation. Con A and LCA, both of which recognize the same specific sugars (mannose, glucose), displayed only a moderate blocking effect. Unlike the native lectin, succinylated WGA, which has the ability to bind to N-acetylglucosamine but not to sialic acid residues, and other lectins examined in this study did not inhibit the action of NGF on Nsp100. WGA-mediated inhibition of NGF action was reversed by the addition of N-acetylglucosamine and by the addition of a much lower concentration of a sialoglycoprotein, mucin, into the culture. Since the binding of succinylated WGA to N-acetylglucosamine residues of cell-surface glycoconjugates is not sufficient to prevent the action of NGF, WGA might act on sialic acid residues of the NGF receptor molecule to effect the inhibition of biological actions of NGF.
Hashimoto
S
S
Department of Biochemistry, Tohoku Dental University, Koriyama, Japan.
Hagino
A
A
D000310:Adrenal Gland Neoplasms / Q000378:metabolism; D000818:Animals; D003208:Concanavalin A / Q000494:pharmacology*; D037102:Lectins / Q000494:pharmacology*; D009414:Nerve Growth Factors / Q000378:metabolism*; D010673:Pheochromocytoma / Q000378:metabolism; D010766:Phosphorylation; D037121:Plant Lectins; D011494:Protein Kinases / Q000378:metabolism*; D051381:Rats; D014407:Tumor Cells, Cultured / Q000378:metabolism; D014909:Wheat Germ Agglutinins / Q000494:pharmacology*
D037102:Lectins; D009414:Nerve Growth Factors; D037121:Plant Lectins; D014909:Wheat Germ Agglutinins; C033619:lentil lectin; D003208:Concanavalin A; D011494:Protein Kinases; C046342:Nsp100 kinase