A pseudosubstrate peptide inhibits protein kinase C-mediated phosphorylation in permeabilized Rat-1 cells.
Activation of protein kinase C (PKC) in Rat-1 fibroblasts leads to rapid phosphorylation of an 80-kDa protein, a major substrate of PKC. Digitonin-permeabilized cells perfectly supported this early response. Introduction of a PKC pseudosubstrate peptide inhibited 80 kDa phosphorylation with an IC50 of 1 microM, while a control peptide had no effect. The results indicate that this semi-intact cell system can be used in combination with the inhibitory pseudosubstrate peptide to study the involvement of PKC in cellular processes.
Eichholtz
T
T
Department of Cellular Biochemistry, The Netherlands Cancer Institute, Amsterdam.
Alblas
J
J
van Overveld
M
M
Moolenaar
W
W
Ploegh
H
H
D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002463:Cell Membrane Permeability / Q000187:drug effects; D004072:Digitonin / Q000494:pharmacology; D015180:Electrophoresis, Gel, Two-Dimensional; D005347:Fibroblasts / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000494:pharmacology*; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors* / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D010455:Peptides; D010750:Phosphoproteins; D000255:Adenosine Triphosphate; D011493:Protein Kinase C; D004072:Digitonin; D013755:Tetradecanoylphorbol Acetate