title: Fluorescent nanoparticles consisting of lipopeptides and fluorescein-modified polyanions for monitoring of protein kinase activity.
abstract: Protein kinase (PK)-responsive nanoparticles (NPs) comprising a hydrophobically modified peptide substrate for PKs and a fluorescein-labeled polyanion (pA-F) were reported for monitoring PK activity via fluorescence intensity measurements. In this system, the formation of NPs by mixing lipopeptides and pA-Fs results in fluorescence quenching, while the quenched fluorescence recovered following dissociation of the NPs owing to the phosphorylation reaction of PKs. Eleven lipopeptides with different hydrophobic moieties (hydrocarbon and lithocholic acid) and four pA-Fs having main chains with differing flexibilities and fluorescein contents were synthesized and used to fabricate a series of twenty-four PK-responsive NP probes. The responses of the PK-responsive NP probes to PKs were evaluated to screen the most suitable NP probes. The assay system was then used to determine the IC(50) values for five inhibitors, the results of which were very similar to those previously reported. Thus, PK-responsive NPs are useful tools for high-throughput screening (HTS) of PK inhibitors.
authors: Graduate School of Systems Life Sciences, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka 819-0395, Japan.
authors: Haruka
authors: H
authors: Koga
authors: Riki
authors: R
authors: Toita
authors: Takeshi
authors: T
authors: Mori
authors: Tetsuro
authors: T
authors: Tomiyama
authors: Jeong-Hun
authors: JH
authors: Kang
authors: Takuro
authors: T
authors: Niidome
authors: Yoshiki
authors: Y
authors: Katayama
mesh_terms: D019793:Fluorescein; D005453:Fluorescence; D057166:High-Throughput Screening Assays / Q000379:methods*; D020128:Inhibitory Concentration 50; D055666:Lipopeptides; D015335:Molecular Probes / Q000138:chemical synthesis; D053758:Nanoparticles / Q000737:chemistry; D000071228:Polyelectrolytes; D011108:Polymers; D047428:Protein Kinase Inhibitors / Q000138:chemical synthesis; D011494:Protein Kinases / Q000032:analysis / Q000378:metabolism*
chemical_list: D019793:Fluorescein; D055666:Lipopeptides; D015335:Molecular Probes; D000071228:Polyelectrolytes; D011108:Polymers; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases