Interaction of alcohols and anesthetics with protein kinase Calpha.
The key signal transduction enzyme protein kinase C (PKC) contains a hydrophobic binding site for alcohols and anesthetics (Slater, S. J., Cox, K. J. A., Lombardi, J. V., Ho, C., Kelly, M. B., Rubin, E., and Stubbs, C. D. (1993) Nature 364, 82-84). In this study, we show that interaction of n-alkanols and general anesthetics with PKCalpha results in dramatically different effects on membrane-associated compared with lipid-independent enzyme activity. Furthermore, the effects on membrane-associated PKCalpha differ markedly depending on whether activity is induced by diacylglycerol or phorbol ester and also on n-alkanol chain length. PKCalpha contains two distinct phorbol ester binding regions of low and high affinity for the activator, respectively (Slater, S. J., Ho, C., Kelly, M. B., Larkin, J. D., Taddeo, F. J., Yeager, M. D., and Stubbs, C. D. (1996) J. Biol. Chem. 271, 4627-4631). Short chain n-alkanols competed for low affinity phorbol ester binding to the enzyme, resulting in reduced enzyme activity, whereas high affinity phorbol ester binding was unaffected. Long chain n-alkanols not only competed for low affinity phorbol ester binding but also enhanced high affinity phorbol ester binding. Furthermore, long chain n-alkanols enhanced phorbol ester induced PKCalpha activity. This effect of long chain n-alkanols was similar to that of diacylglycerol, although the n-alkanols alone were weak activators of the enzyme. The cellular effects of n-alkanols and general anesthetics on PKC-mediated processes will therefore depend in a complex manner on the locality of the enzyme (e.g. cytoskeletal or membrane-associated) and activator type, apart from any isoform-specific differences. Furthermore, effects mediated by interaction with the region on the enzyme possessing low affinity for phorbol esters represent a novel mechanism for the regulation of PKC activity.
Slater
S J
SJ
Department of Pathology and Cell Biology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Kelly
M B
MB
Larkin
J D
JD
Ho
C
C
Mazurek
A
A
Taddeo
F J
FJ
Yeager
M D
MD
Stubbs
C D
CD
D000438:Alcohols / Q000737:chemistry*; D018681:Anesthetics, General / Q000737:chemistry*; D000818:Animals; D002417:Cattle; D007527:Isoenzymes / Q000737:chemistry*; D008024:Ligands; D010703:Phorbol Esters / Q000737:chemistry; D011485:Protein Binding; D011493:Protein Kinase C / Q000737:chemistry*; D051571:Protein Kinase C-alpha; D011994:Recombinant Proteins; D012995:Solubility; D013329:Structure-Activity Relationship
D000438:Alcohols; D018681:Anesthetics, General; D007527:Isoenzymes; D008024:Ligands; D010703:Phorbol Esters; D011994:Recombinant Proteins; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha