Synthesis Of Nanomaterials And Their Applications In Detecting Of Kinase Activity

This thesis included a review and the research work. In the review section, theintroduction in chapter1provides series of concepts, various characteristics and applicationsof nanomaterials in analytical chemistry. And then we introduced the physiologicalsignificance of protein kinase, and a general introduction was proposed about the methods ofanalysis for the detection of protein kinase activities. Finally, we summarize the applicationsof different nanomaterials in detecting of kinase activity.The research work contained the following subunits：1. A simple and sensitive photoluminescence (PL) assay for the activity of a proteinkinase CK2based on the selective aggregation of phosphorylated peptide graphene quantumdot (GQD) conjugates triggered by Zr4+ion coordination has been established. Under optimalconditions, a linear relationship between the decreased PL intensity of peptide GQDconjugates and the concentration of casein kinase II (CK2) was obtained. The proposedmethod showed potential applications in kinase inhibitor screening. To demonstrate thepotential of this GQD-based platform for screening of kinase inhibitors in real biologicalsystems, the inhibition of CK2phosphorylation activity by four different inhibitors (ellagicacid,5,6-dichlorobenzimidazole-1-β-D-ribofuranoside, emodin, and quercetin) was tested inhuman serum by comparing signals from samples incubated with the inhibitors against thatwithout any inhibitor.2. We present a novel method to detect the activity of a protein kinase PKA by theselective aggregation of phosphorylated peptide-Au nanoparticles (AuNPs) conjugatestriggered by Zr4+ion coordination. With more sophisticated design of the peptide substratesequences, detecting other enzymes could also be possible. Under optimal conditions, withthe increase of the concentration of PKA, the UV absorption intensity decreased, whichindicated the aggregation of phosphorylated peptide-AuNPs by Zr4+ion. What’s more, theproposed method showed potential applications in kinase PKA inhibitor screening. Takeellagic acid that the inhibitor of PKA as an example, the multicoordinative interactionsbetween phosphorylated peptide-AuNPs and Zr4+ion causes the aggregation, which inducesthe increase of the intensity of light scattering. We can obtain the IC50that the inhibitorconcentration producing50%inhibition for ellagic acid.