A light-activated probe of intracellular protein kinase activity.
The first example of a photoactivated probe of intracellular enzymatic activity is described. The caged derivative of a fluorescent protein kinase C peptide-based sensor was prepared by modifying the free hydroxyl group of a phosphorylatable serine moiety with a photolabile appendage that blocks phosphoryl transfer. We have demonstrated that the caged sensor allows one to (1) sample PKC activity with exquisite temporal precision, (2) control the relative amount of active sensor available for phosphorylation, and (3) examine protein kinase activity at multiple time points.
Veldhuyzen
Willem F
WF
Department of Biochemistry, The Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, New York 10461, USA.
Nguyen
Quan
Q
McMaster
Gary
G
Lawrence
David S
DS
D000081:Acetamides; D000596:Amino Acids / Q000737:chemistry; D001120:Arginine / Q000737:chemistry; D015374:Biosensing Techniques / Q000379:methods; D062845:Chloroacetates; D002851:Chromatography, High Pressure Liquid; D005449:Fluorenes / Q000737:chemistry; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D010782:Photolysis; D011493:Protein Kinase C / Q000737:chemistry / Q000378:metabolism*; D014238:Trichloroacetic Acid / Q000737:chemistry
D000081:Acetamides; D000596:Amino Acids; D062845:Chloroacetates; D005449:Fluorenes; D005456:Fluorescent Dyes; C016456:N(alpha)-fluorenylmethyloxycarbonylamino acids; D014238:Trichloroacetic Acid; D001120:Arginine; D011493:Protein Kinase C; C009528:trichloroacetamide