Calmodulin-dependent protein kinases in rat glioblastoma.
The mitogenic activity of several growth factors is mediated by calcium-dependent signal transduction. Calmodulin (CaM) binding proteins such as CaM-dependent protein kinases are important components of this pathway and may be altered in diseases characterized by abnormal cell growth. CaM kinase II is believed to regulate the phosphorylation of microtubular-associated proteins and control the initiation of DNA synthesis. Furthermore, drugs that inhibit CaM-mediated signal transduction also inhibit cellular proliferation and are cytotoxic to numerous malignant cell lines, including those established from malignant gliomas. Yet, little is known about CaM-dependent protein kinases in these tumors. Therefore, we have investigated the activity and distribution of CaM-dependent protein kinase II in normal and malignant glial tissues, a kinase believed to play a critical role in cell cycle regulation. C6 and 9L cells contained kinase activities that were activated by Ca2+/CaM and inhibited by trifluoperazine. Tissue extracts from these cell lines and from rat brain white matter phosphorylated exogenous synapsin I in a pattern consistent with the presence of CaM kinase II activity as determined by phosphopeptide mapping. CaM kinase II activity was confirmed using a specific peptide substrate and inhibitor. An unexpected finding was that glioma lines, but not rat brain white matter, also contained a CaM-dependent protein kinase detected by the phosphorylation of a M(r) 100,000 protein, subsequently identified as elongation factor 2, the only known substrate for CaM kinase III.(ABSTRACT TRUNCATED AT 250 WORDS)
Cheng
E H
EH
Gorelick
F S
FS
Czernik
A J
AJ
Bagaglio
D M
DM
Hait
W N
WN
D000818:Animals; D001932:Brain Neoplasms / Q000201:enzymology*; D002118:Calcium / Q000378:metabolism; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000187:drug effects / Q000378:metabolism*; D002147:Calmodulin / Q000378:metabolism; D005909:Glioblastoma / Q000201:enzymology*; D015151:Immunoblotting; D008297:Male; D010449:Peptide Mapping; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction; D016704:Synapsins / Q000378:metabolism; D014268:Trifluoperazine / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D002147:Calmodulin; D016704:Synapsins; D014268:Trifluoperazine; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002118:Calcium