Staurosporine activates a 60,000 M(r) protein kinase in bovine chromaffin cells that phosphorylates myelin basic protein in vitro.
Bovine chromaffin cells contain a family of renaturable protein kinases. One of these, a 60,000 M(r) kinase (PK60) that phosphorylated myelin basic protein in vitro, was activated fourfold when cells were treated with the protein kinase inhibitor staurosporine. Because staurosporine inhibits protein kinase C, the role of this kinase in the regulation of PK60 activity was investigated. Fifty nanomolar staurosporine produced half-maximal inhibition of protein kinase C activity in chromaffin cells, whereas approximately 225 nM staurosporine was required to induce half-maximal activation of PK60. Other protein kinase C inhibitors, H-7 and K-252a, did not mimic the effect of staurosporine on PK60 activity. Chromaffin cells have three protein kinase C isoforms: alpha, epsilon, and zeta. Prolonged treatment with phorbol esters depleted the cells of protein kinase C alpha and epsilon, but not zeta. Neither activation nor depletion of protein kinase C affected the basal activity of PK60. Moreover, staurosporine activated PK60 in cells depleted of protein kinase C alpha and epsilon; thus, staurosporine appeared to activate PK60 by a mechanism that does not require these protein kinase C isoforms. Incubation of cell extracts with staurosporine in vitro did not activate PK60. Incubation of these extracts with adenosine 5'-O-(3-thiotriphosphate), however, caused a twofold activation of PK60. Although this suggests that PK60 activity is regulated by phosphorylation, the mechanism by which staurosporine activates PK60 is not known. Staurosporine has been reported to promote neurite outgrowth from chromaffin cells. The role of PK60 in mediating the effects of staurosporine on chromaffin cell function remains to be determined.
Pavlović-Surjancev
B
B
Department of Pediatrics, University of Chicago, Illinois 60637.
Cahill
A L
AL
Perlman
R L
RL
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D000311:Adrenal Glands / Q000187:drug effects / Q000201:enzymology; D000470:Alkaloids / Q000494:pharmacology*; D000818:Animals; D002417:Cattle; D002838:Chromaffin System / Q000187:drug effects / Q000201:enzymology*; D004789:Enzyme Activation / Q000187:drug effects; D015151:Immunoblotting; D007700:Kinetics; D008970:Molecular Weight; D004676:Myelin Basic Protein / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000470:Alkaloids; D004676:Myelin Basic Protein; C022571:adenosine 5'-O-(3-thiotriphosphate); D000255:Adenosine Triphosphate; D011494:Protein Kinases; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate