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Retinola induced changes in cAMP‐dependent protein kinase activity detected by a new minigel assay
Author(s) -
Hohmann Philip,
Greene Robert S.
Publication year - 1990
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(90)80641-u
Subject(s) - kinase , gene isoform , protein kinase a , biochemistry , chemistry , peptide , enzyme , cell culture , substrate (aquarium) , in vitro , microbiology and biotechnology , biology , ecology , genetics , gene
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 10Tcells. Protein kinases in 0.5 μl aliquots of cell extracts were resolved on nondenaturing gradient Phastgels and assayed for PKA activity in situ using a specific peptide substrate and [ 32 P]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.