Retinoid induced changes in cAMP-dependent protein kinase activity detected by a new minigel assay.
A new method for the resolution of protein kinases in nondenaturing minigels was used to analyze the type of PKA induced by retinyl acetate in transformed mouse 10T1/2 cells. Protein kinases in 0.5 microliters aliquots of cell extracts were resolved on nondenaturing gradient Phastgels and assayed for PKA activity in situ using a specific peptide substrate and [32P]ATP. Retinyl acetate caused a decrease in type I and an increase in type II PKA isoforms after about 36 h in culture. The system provides a quick and sensitive means for analyzing the activity of PKA isoforms from small numbers of cells.
Hohmann
P
P
Department of Experimental Biology, Roswell Park Cancer Institute, Buffalo, NY 14263.
Greene
R S
RS
D000818:Animals; D002461:Cell Line, Transformed; D000242:Cyclic AMP / Q000494:pharmacology*; D004224:Diterpenes; D004591:Electrophoresis, Polyacrylamide Gel; D004790:Enzyme Induction / Q000187:drug effects; D007527:Isoenzymes / Q000096:biosynthesis* / Q000302:isolation & purification; D051379:Mice; D011494:Protein Kinases / Q000096:biosynthesis* / Q000302:isolation & purification; D000084562:Retinyl Esters; D014801:Vitamin A / Q000031:analogs & derivatives* / Q000494:pharmacology
D004224:Diterpenes; D007527:Isoenzymes; D000084562:Retinyl Esters; D014801:Vitamin A; C009166:retinol acetate; D000242:Cyclic AMP; D011494:Protein Kinases