Fluorescent polyion complex nanoparticle that incorporates an internal standard for quantitative analysis of protein kinase activity.
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.
Department of Applied Chemistry, Faculty of Engineering, ‡Graduate School of Systems Life Sciences, §Center for Future Chemistry, ||International Research Center for Molecular Systems, ⊥Center for Advanced Medical Innovation, and #Innovation Center for Medical Redox Navigation, Kyushu University , 744 Motooka, Nishi-ku, Fukuoka 819-0395, Japan.
Takanobu
T
Nobori
Shujiro
S
Shiosaki
Takeshi
T
Mori
Riki
R
Toita
Chan Woo
CW
Kim
Yuta
Y
Nakamura
Akihiro
A
Kishimura
Takuro
T
Niidome
Yoshiki
Y
Katayama
D055162:Biocatalysis; D002232:Carbocyanines / Q000737:chemistry* / Q000378:metabolism; D004789:Enzyme Activation; D057075:Enzyme Assays / Q000379:methods*; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism; D007477:Ions / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D015394:Molecular Structure; D053758:Nanoparticles / Q000737:chemistry* / Q000378:metabolism; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D010766:Phosphorylation; D051571:Protein Kinase C-alpha / Q000032:analysis / Q000378:metabolism*; D012015:Reference Standards; D012235:Rhodamines / Q000737:chemistry* / Q000378:metabolism
D002232:Carbocyanines; D005456:Fluorescent Dyes; D007477:Ions; D010455:Peptides; D012235:Rhodamines; C085321:cyanine dye 5; C017645:polyaspartate; C005358:tetramethylrhodamine; D051571:Protein Kinase C-alpha