NGF stimulates incorporation of fucose or glucosamine into an external glycoprotein in cultured rat PC12 pheochromocytoma cells

doi:10.1016/0092-8674(78)90004-1

Cell

Volume 15, Issue 2, October 1978, Pages 357-365

https://doi.org/10.1016/0092-8674(78)90004-1 Get rights and content

Abstract

Rat PC12 pheochromocytoma cells respond to Nerve Growth Factor (NGF) by ceasing to undergo mitosis and acquiring neuronal characteristics, including outgrowth of neurites and electrical excitability. We have found that NGF treatment results in no consistent qualitative and only a few minor quantitative changes in the two-dimensional electrophoresis pattern of ¹⁴C-amino acid-labeled proteins synthesized by the cells. On the other hand, NGF stimulates the incorporation of radiolabeled fucose or glucosamine into several components, including one of apparent molecular weight 230,000 daltons on one- and two-dimensional SDS-polyacrylamide gels. This component is removed by mild trypsinization of intact cells and therefore appears to be a glycoprotein (named here NILE) at least partially exposed on the cell surface. Stimulation of NILE glycoprotein labeling can first be detected after 2 days of exposure to NGF and increases progressively with time of treatment. This change is not solely a consequence of the cessation of cell division caused by NGF, since it does not occur in nondividing PC12 populations prepared by cytosine arabinoside treatment. NGF stimulates labeling of NILE glycoprotein even when attachment and process outgrowth are prevented by growing the cells in spinner suspension cultures. The relative rate of labeling attained under these conditions is less, however, than in NGF-treated, substrate-attached cells. Stimulation of NILE glycoprotein labeling by NGF is selectively blocked (as is neurite outgrowth) by camptothecin, an RNA synthesis inhibitor, and thus may require transcription. Despite their similarities in apparent size and exposure on cell surfaces, the NILE and LETS glycoproteins are shown to be immunologically distinct.

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This chapter reviews some of the structural features of the immunoglobulin (Ig)-CAMS. It describes some of their interactions with other molecules of the same and different types and discusses some aspects of the intracellular signalling mediated through this binding. One of the distinguishing characteristics of immunoglobins and Ig-like molecules is a protein structure comprised of multiple domains. All of the neural Ig-CAMS are glycoproteins and subject to posttranslational modification. Neural Ig-CAMS can also be phosphorylated, almost always on serine or threonine. The synthesis and processing of Ig-CAMS can be very complex. The members of the neuronal Ig superfamily can be distinguished and classified in a number of ways. The data on the binding of Ig-CAMS to intracellular elements is rather sparse, but because such an interaction would provide a mechanism for communicating signals from outside to inside the cell or vice versa, it remains an attractive hypothesis.

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Cell

ArticleNGF stimulates incorporation of fucose or glucosamine into an external glycoprotein in cultured rat PC12 pheochromocytoma cells

Abstract

Cell

J. Biol. Chem.

Cell Differentiation

Arch. Biochem. Biophys.

J. Biol. Chem.

Cell

Brain Res.

Biochim. Biophys. Acta

Cell

J. Biol. Chem.

Int. J. Biochem.

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

Brain Res.

Cell

J. Mol. Biol.

J. Biol. Chem.

J. Biol. Chem.

Three abundance classes in HeLa cell messenger RNA

Nature

The nerve growth factor: purification as a 30, 000-molecular-weight protein

A film detection method for tritium-labeled proteins and nucleic acids in polyacrylamide gels

Eur. J. Biochem.

Proteolytic enzymes initiating cell division and escape from contact inhibition of growth

Nature

Article
NGF stimulates incorporation of fucose or glucosamine into an external glycoprotein in cultured rat PC12 pheochromocytoma cells