1. Fluorescent Polyion Complex Nanoparticle That Incorporates an Internal Standard for Quantitative Analysis of Protein Kinase Activity
- Author
-
Takanobu Nobori, Riki Toita, Yuta Nakamura, Shujiro Shiosaki, Chan Woo Kim, Takeshi Mori, Yoshiki Katayama, Takuro Niidome, and Akihiro Kishimura
- Subjects
Models, Molecular ,Protein Kinase C-alpha ,Fluorophore ,Biomedical Engineering ,Pharmaceutical Science ,Nanoparticle ,Bioengineering ,Fluorescence ,chemistry.chemical_compound ,Phosphorylation ,Protein kinase A ,Enzyme Assays ,Fluorescent Dyes ,Ions ,Pharmacology ,Molecular Structure ,Rhodamines ,Organic Chemistry ,Substrate (chemistry) ,Carbocyanines ,Reference Standards ,Enzyme Activation ,chemistry ,Biochemistry ,Biocatalysis ,Nanoparticles ,Peptides ,Quantitative analysis (chemistry) ,Biotechnology - Abstract
We demonstrate a polyion complex (PIC) nanoparticle that contains both a responsive fluorophore and an "internal standard" fluorophore for quantitative measurement of protein kinase (PK) activity. The PK-responsive fluorophore becomes more fluorescent with PK-catalyzed phosphorylation of substrate peptides incorporated in the PIC, while fluorescence from the internal standard remains unchanged during phosphorylation. This new concept will be useful for quantitative PK assays and the discovery of PK inhibitors.
- Published
- 2014