Kinase-Catalyzed Modification of Gold Nanoparticles:  A New Approach to Colorimetric Kinase Activity Screening

Abstract

Peptide-stabilized gold nanoparticles have been enzymatically biotinylated by a kinase-catalyzed reaction using biotin-ATP as a cosubstrate. Upon mixing with avidin-modified particles, solutions of biotinylated particles change color from red to blue, indicating aggregation of particles. On the basis of this reaction, we have developed a simple colorimetric test to monitor kinase inhibitor activity.

Applications of Nanoparticles in Biology

“…[70] In presence of thermolysin the peptides are fragmented and the assembly changes color from blue to red with a sensitivity of 90 zg mL À1 (i.e., less than 380 molecules of protease). [71] This approach has been extended to kinases [72] and phosphatases [73,74] at low concentration.…”

Gold-nanoparticle-based biosensors for detection of enzyme activity

“…Over the last few years, a great variety of morphologically different GNPs have been prepared, such as rods, urchins, prisms, wires, disks, and stars, with varying optical properties according to their size, shape, and surface chemistry [32]. In contrast to the well-studied and most frequently used spherical particles, many of these other-shaped particles have optical resonances in the nearinfrared spectral range, allowing deeper penetration within tissues and an absence of photochemical damage; these features are highly advantageous for intracellular or in vivo imaging applications.…”

Gold nanoparticles-based electrochemical method for the detection of protein kinase with a peptide-like inhibitor as the bioreceptor

“…51,52 Among these methods, colorimetric assays based on the aggregation or redispersion of gold nanoparticles (AuNPs) are in particular prevalent because of their simple manipulation principle and easy detection procedure. Usually, there are two categories of the colorimetric sensors based on the AuNPs used: one is based on the cross-linking aggregation of AuNPs functionalized respectively with recognition elements (eg, avidin and antibody) and kinase-specific peptide, 10,11,14,15 and the other is based on the non-cross-linking aggregation of unmodified AuNPs, which is tuned by the phosphorylation-induced net charge change of substrate peptide. 12,53,54 Although the cross-linking strategies are robust and highly specific, they suffer from the complicated functionalization of AuNPs.…”