Measurement of kinase activation in single mammalian cells.
We demonstrate a new method for the simultaneous measurement of the activation of key regulatory enzymes within single cells. To illustrate the capabilities of the technique, the activation of protein kinase C (PKC), protein kinase A (PKA), calcium-calmodulin activated kinase II (CamKII), and cdc2 protein kinase (cdc2K) was measured in response to both pharmacological or physiological stimuli. This assay strategy should be applicable to a broad range of intracellular enzymes, including phosphatases, proteases, nucleases, and other kinases.
Meredith
G D
GD
Department of Physiology and Biophysics, University of California, Irvine, CA 92697-4560, USA.
Sims
C E
CE
Soughayer
J S
JS
Allbritton
N L
NL
D016475:3T3 Cells; D000818:Animals; D001671:Biochemistry / Q000379:methods*; D016203:CDC2 Protein Kinase / Q000096:biosynthesis; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000096:biosynthesis; D017868:Cyclic AMP-Dependent Protein Kinases / Q000096:biosynthesis; D019075:Electrophoresis, Capillary / Q000379:methods; D004789:Enzyme Activation; D051379:Mice; D010455:Peptides; D010770:Phosphotransferases / Q000096:biosynthesis*; D011493:Protein Kinase C / Q000096:biosynthesis; D051381:Rats; D015398:Signal Transduction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013997:Time Factors; D014407:Tumor Cells, Cultured
D010455:Peptides; D010770:Phosphotransferases; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D016203:CDC2 Protein Kinase; D013755:Tetradecanoylphorbol Acetate