Fibroblast growth factor-induced decrease in the phosphorylation of Nsp100 mediated through a calcium-dependent mechanism and blocked by lectins.
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概要
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Separate treatment of PC12h cells with basic fibroblast growth factor (bFGF) and with epidermal growth factor (EGF) induced a selective decrease in the incorporation of radioactive phosphate into a 100, 000-dalton soluble protein during phosphorylation with (γ-32P)ATP of soluble extracts from the cells, as was seen previously with nerve growth factor (NGF). This 100, 000-dalton soluble protein was designated in earlier studies as nerve growth factor-sensitive protein 100 (Nsp 100). The inhibitory effects of bFGF and EGF on Nsp 100 phosphorylation were prevented by pretreatment of PC12h cells with the calcium chelator, EGTA. Treatment of PC12h cells with the plant lectin wheat germ agglutinin (WGA), which binds to N-acetylglucosamine and sialic acid residues on glycoconjugates, blocked the inhibitory effects of bFGF, EGF, and NGF on Nsp 100 phosphorylation. The blockage by WGA was reversed by the addition of the lectin-specific sugar N-acetylglucosamine to the PC12h cultures. Although pretreatment of PC12h cells with succinylated WGA, which has the ability to bind to N-acetylglucosamine but not to sialic acid residues, failed to block the inhibitory effect of NGF on Nsp 100 phosphorylation as described previously, it did prevent the inhibitory effect of bFGF on this phosphorylation. These data suggest that in PC12h cells bFGF and EGF induce a decrease in the phosphorylation of Nsp 100 mediated through a Ca2+- dependent mechanism, as in the case of NGF. Furthermore, the blockage of the bFGF-induced inhibition of Nsp 100 phosphorylation by WGAand its succinylated form indicates that N-acetylglucosamine residues of bFGF receptor molecules might be involved in the mechanism by which bFGF inhibits the phosphorylation. On the other hand, staurosporine, a protein kinase inhibitor, blocked the NGF-induced decreGase in the phosphorylation of Nspl00, but failed to block the bFGF-induced one.
- 日本細胞生物学会の論文
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