Specificity of action of bisindolylmaleimide protein kinase C inhibitors: do they inhibit the 70kDa ribosomal S6 kinase in cardiac myocytes?
Bisindolylmaleimide protein kinase C (PKC) inhibitors, such as GF109203X and Ro31-8220, are used as pharmacological tools in many cellular systems. However, in vitro, GF109203X and Ro31-8220 also inhibit the 70kDa ribosomal S6 kinase (p70(S6K)) with similar potency. We determined whether GF109203X and Ro31-8220 inhibit p70(S6K) activity in intact adult rat ventricular myocytes (ARVM). First, we confirmed that increased phosphorylation of the 40S ribosomal S6 protein (a cellular substrate for both p70(S6K) and the 90 kDa ribosomal S6 kinase) in response to stimulation of ARVM by insulin-like growth factor-1 (300 ng/mL; 10 min) occurs specifically through rapamycin-sensitive activation of p70(S6K). Then, using this response as the index of cellular p70(S6K) activity, we determined the effects of GF109203X and Ro31-8220 (1, 3 or 10 microM) on such activity. At these concentrations, neither GF109203X nor Ro31-8220 inhibited cellular p70(S6K) activity. In contrast, even at 1 microM, cellular PKC activity (stimulated by a 3 min exposure to 30 nM phorbol 12-myristate 13-acetate) was significantly inhibited by each agent. We conclude that; (1) data obtained in vitro may not necessarily be extrapolated to intact cells and (2) inhibition of p70(S6K) is unlikely to contribute to the actions of GF109203X and Ro31-8220 in ARVM.
Cardiovascular Division, King's College London, The Rayne Institute, St Thomas' Hospital, London SE17EH, UK.
Neil A
NA
Roberts
Michael S
MS
Marber
Metin
M
Avkiran
D000818:Animals; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology*; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology; D008297:Male; D008301:Maleimides / Q000494:pharmacology*; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000201:enzymology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors*; D051381:Rats; D017208:Rats, Wistar; D038601:Ribosomal Protein S6 / Q000378:metabolism; D038762:Ribosomal Protein S6 Kinases, 70-kDa / Q000037:antagonists & inhibitors* / Q000378:metabolism
D004791:Enzyme Inhibitors; D007211:Indoles; D008301:Maleimides; D038601:Ribosomal Protein S6; D007334:Insulin-Like Growth Factor I; D038762:Ribosomal Protein S6 Kinases, 70-kDa; D011493:Protein Kinase C; C088060:bisindolylmaleimide; C064758:Ro 31-8220