Measurement of kinase activation in single mammalian cells

Nat Biotechnol. 2000 Mar;18(3):309-12. doi: 10.1038/73760.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 3T3 Cells
  • Animals
  • Biochemistry / methods*
  • CDC2 Protein Kinase / biosynthesis
  • Calcium-Calmodulin-Dependent Protein Kinases / biosynthesis
  • Cyclic AMP-Dependent Protein Kinases / biosynthesis
  • Electrophoresis, Capillary / methods
  • Enzyme Activation*
  • Mice
  • Peptides
  • Phosphotransferases / biosynthesis*
  • Protein Kinase C / biosynthesis
  • Rats
  • Signal Transduction
  • Tetradecanoylphorbol Acetate / pharmacology
  • Time Factors
  • Tumor Cells, Cultured

Substances

  • Peptides
  • Phosphotransferases
  • Cyclic AMP-Dependent Protein Kinases
  • Protein Kinase C
  • Calcium-Calmodulin-Dependent Protein Kinases
  • CDC2 Protein Kinase
  • Tetradecanoylphorbol Acetate