Differential tyrosine phosphorylation/activation of oncogenic proline-directed protein kinase F(A)/GSK-3alpha in well and poorly differentiated human prostate carcinoma cells.

Abstract

Computer analysis of protein phosphorylation site sequences revealed that transcriptional factors and viral oncoproteins are prime targets for regulation of proline-directed protein phosphorylation, suggesting an association of the proline-directed protein kinase (PDPK) family with neoplastic transformation and tumorigenesis. In this report, an immunoprecipitate activity assay of proline-directed protein kinase F(A)/glycogen synthase kinase-3alpha (PDPK F(A)/GSK-3alpha) has been optimized to demonstrate significantly increased (p < 0.01) activity in poorly differentiated human prostate carcinoma PC-3 cells (55.5+/-3.8 units/mg) when compared to well-differentiated LNCaP cells (28.1+/-2.3 units/mg). Immunoblotting analysis revealed that increased activity of this PDPK in PC-3 cells is due not to overexpression of the protein, but to enhanced tyrosine phosphorylation of the kinase. When treated with genistein (a protein tyrosine kinase PTK inhibitor), the enhanced tyrosine phosphorylation/activation of the kinase in PC-3 cells can be blocked. Conversely, when treated with vanadate (a protein tyrosine phosphatase PTP inhibitor), the phosphotyrosine content of PDPK F(A)/GSK-3alpha in LNCaP cells can be promoted to the level of PC-3 cells. In sharp contrast, the PTK inhibitor has little effect on the tyrosine phosphorylation level of the kinase in LNCaP cells, whereas the PTP inhibitor has little effect on the tyrosine phosphorylation level of the kinase in PC-3 cells. Taken together, the results provide initial evidence that the tyrosine phosphorylation/activation levels of this oncogenic PDPK can be differentially regulated in well- and poorly differentiated prostate carcinoma cells.

• Authors: C C Yang, C P Hsu, J C Sheu, X Y Mai, S D Yang
• Journal: Journal of protein chemistry (J Protein Chem) Vol. 17 Issue 4 Pg. 329-35 (May 1998) ISSN: 0277-8033 [Print] United States
• PMID: 9619586 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Enzyme Inhibitors
• Vanadates
• Tyrosine
• Genistein
• Calcium-Calmodulin-Dependent Protein Kinases
• Glycogen Synthase Kinase 3

Topics

• Amino Acid Sequence
• Calcium-Calmodulin-Dependent Protein Kinases (antagonists & inhibitors, metabolism)
• Cell Differentiation
• Enzyme Activation
• Enzyme Inhibitors (pharmacology)
• Genistein (pharmacology)
• Glycogen Synthase Kinase 3
• Humans
• Male
• Molecular Sequence Data
• Oncogenes
• Phosphorylation
• Prostatic Neoplasms (enzymology, pathology)
• Tumor Cells, Cultured
• Tyrosine (metabolism)
• Vanadates (pharmacology)