Effects of 6- and 8-substituted analogs of adenosine 3':5'-monophosphate on phosphoenolpyruvate carboxykinase and tyrosine aminotransferase in hepatoma cell cultures.
A variety of 6- and 8-substituted analogs of cAMP (cyclic adenosine 3:5-monophosphate) have been tested for their ability to increase activity of tyrosine aminotransferase (EC 2.6.1.5) in cultured Reuber H35 hepatoma cells. Some analogs, particularly the 8-thio-substituted ones, produced effects approximately equivalent to those generated by N-6, O2'-dibutyryl cAMP. In contrast, cAMP and its O-2-monobutyryl derivative were relatively ineffective even at very high concentrations, whereas three other analogs actually depressed the activity of the aminotransferase. Changes in enzyme activity generated by the various analogs were paralleled closely by changes in the relative rate of aminotransferase synthesis. An excellent correlation was found to exist between the ability of any given analog to influence the activity of tyrosine aminotransferase and that of phosphoenolpyruvate carboxykinase (EC 4.1.1.32). A similar correlation was found to exist between the ability of various analogs to evelate the activity of these enzymes and to inhibit reversibly the growth of H35 cells. Only one of five inhibitors of cAMP phosphodiesterase activity tested produce any increase in aminotransferase activity when added alone. All of the 6- and 8-substituted analogs tested, including noniducers, stimulated f1 histone phosphorylation in crude rat liver extracts with approximately equal potencies. On the other hand, dibutyryl cAMP was only a weak activator of protein kinase in vitro, even though it is a potent enzyme inducer. A possible resolution of this apparent discrepancy has been provided by preliminary analyses of site-specific f1 histone phosphorylation in whole cells. Only compounds active as aminotransferase inducers are capable of stimulating phosphorylation of the serine-37 residue of endogenous f1 histone (3- to 10-fold).
D000818:Animals; D002087:Butyrates; D002250:Carbon Radioisotopes; D006528:Carcinoma, Hepatocellular / Q000201:enzymology*; D002460:Cell Line; D000242:Cyclic AMP / Q000031:analogs & derivatives* / Q000494:pharmacology; D006152:Cyclic GMP / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D007292:Inosine Nucleotides / Q000494:pharmacology; D007700:Kinetics; D008113:Liver Neoplasms; D009374:Neoplasms, Experimental / Q000201:enzymology; D009712:Nucleotides, Cyclic / Q000494:pharmacology; D010726:Phosphodiesterase Inhibitors; D010729:Phosphoenolpyruvate Carboxykinase (GTP) / Q000378:metabolism*; D010761:Phosphorus Radioisotopes; D011494:Protein Kinases / Q000378:metabolism; D051381:Rats; D013329:Structure-Activity Relationship; D013806:Theophylline / Q000494:pharmacology; D013997:Time Factors; D014316:Tritium; D014444:Tyrosine Transaminase / Q000378:metabolism*
D002087:Butyrates; D002250:Carbon Radioisotopes; D007292:Inosine Nucleotides; D009712:Nucleotides, Cyclic; D010726:Phosphodiesterase Inhibitors; D010761:Phosphorus Radioisotopes; D014316:Tritium; D013806:Theophylline; D000242:Cyclic AMP; D014444:Tyrosine Transaminase; D011494:Protein Kinases; D010729:Phosphoenolpyruvate Carboxykinase (GTP); D006152:Cyclic GMP