A chromism-based assay (CHROBA) technique for in situ detection of protein kinase activity.
A unique chromism-based assay technique (CHROBA) using photochromic spiropyran-containing peptides has been firstly established for detection of protein kinase A-catalyzed phosphorylation. The alternative method has advantages that avoid isolation and/or immobilization of kinase substrates to remove excess reagents including nonreactive isotope-labeled ATP or fluorescently-labeled anti-phosphoamino acid antibodies from the reaction mixture. Such a novel protocol based on thermocoloration of the spiropyran moiety in the peptide can offer not only an efficient screening method of potent kinase substrates but also a versatile analytical tool for monitoring other post-translational modification activities.
Tomizaki
Kin-ya
KY
Department of Bioengineering and the COE21 Program, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, B-40, 4259 Nagatsuta, Midori, Yokohama 226-8501, Japan.
Jie
Xu
X
Mihara
Hisakazu
H
D001578:Benzopyrans / Q000737:chemistry; D002863:Chromogenic Compounds / Q000737:chemistry*; D003124:Colorimetry / Q000379:methods*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D007211:Indoles; D009574:Nitro Compounds; D010455:Peptides / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
D001578:Benzopyrans; D002863:Chromogenic Compounds; D007211:Indoles; D009574:Nitro Compounds; D010455:Peptides; C088184:spiropyran; D017868:Cyclic AMP-Dependent Protein Kinases