On-bead fluorescence assay for serine/threonine kinases.
[chemical reaction: see text]. A novel fluorescence-based assay for serine/threonine kinases is described. Base-mediated beta-elimination of the phosphate moiety and the Michael addition of a thiol-containing fluorescent molecule allows convenient and efficient detection of the enzyme activity. This approach may be broadly applicable to various serine/threonine kinases.
Akita
Shoji
S
Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabe-dori, Nagoya, Japan.
Umezawa
Naoki
N
Higuchi
Tsunehiko
T
D000595:Amino Acid Sequence; D047388:Casein Kinases / Q000032:analysis / Q000378:metabolism; D005453:Fluorescence; D005456:Fluorescent Dyes; D015394:Molecular Structure; D010766:Phosphorylation; D011494:Protein Kinases / Q000032:analysis / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000032:analysis* / Q000378:metabolism*; D013379:Substrate Specificity
D005456:Fluorescent Dyes; D011494:Protein Kinases; D047388:Casein Kinases; D017346:Protein Serine-Threonine Kinases