Nerve Growth Factor (NGF) Responses by Non-Neuronal Cells: Detection by Assay of a Novel NGF-Activated Protein Kinase
作者:
VolonteCinzia,
GreeneLloyd A.,
期刊:
Growth Factors
(Taylor Available online 1990)
卷期:
Volume 2,
issue 4
页码: 321-331
ISSN:0897-7194
年代: 1990
DOI:10.3109/08977199009167027
出版商: Taylor&Francis
关键词: nerve growth factor;non-neuronal cells;kinase activity
数据来源: Taylor
摘要:
Past work described the partial purification and characterization of a novel serine protein kinase activity designated protein kinase N (PKN) that is activated by nerve growth factor (NGF) in cultured PC12 cells [Rowland et al. (1987)J. Biol. Chem.262; 7504–7513]. We have now devised a rapid, sensitive technique for partially purifying and assaying PKN activity in cell extracts. This methodology was applied to the IARC-EW-1 osteosarcoma and several additional non-neuronal cell lines that possess NGF receptors but that lack both morphological and a variety of additional biochemical responses to NGF. In each case, NGF significantly elevated PKN activity. The assay also revealed activation of PKN activity in IARC-EW-1 cells by additional agents, including epidermal growth factor, fibroblast growth factor, phorbol ester, and a cAMP analog. Also tested were an NGF-receptor-deficient PC12 cell variant and sublines thereof into which human NGF receptors had been introduced [Hempstead et al. (1989)Science243; 373–375]. Acquisition of the NGF receptors resulted in NGF-activatable PKN activity. These findings indicate that detection of PKN activity may serve as a sensitive means to test NGF responsiveness in cells lacking macroscopic responses to the factor and that non-neuronal cells may be useful for studying primary signaling events in the NGF mechanism of action.
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