A novel label-free electrochemical strategy for monitoring the activity and inhibition of protein kinase is developed, based on the linkage between the phosphorylated peptide and DNA functionalized Au nanoparticles (DNA-AuNPs) by Zr(4+) and the chronocoulometric response of [Ru(NH(3))(6)](3+) absorbed on the DNA-AuNPs.
The sequence-length-dependent adsorption of single-stranded DNA (ssDNA) on unmodified gold nanoparticles is exploited as a new mechanism for colorimetric nuclease assay and measurement of oxidative DNA damage.
The rapid development in nanoparticle production and application during the past decade requires an easy, rapid, and predictive screening method for nanoparticles toxicity assay. In this study, the toxicological effects and the source of toxicity of copper nanoparticles (CuNPs) are investigated based on a stress-responsive bacterial biosensor array. According to the responses of the biosensing strains, it is found that CuNPs induce not only oxidative stress in E. coli, but also protein damage, DNA damage, and cell membrane damage, and ultimately cause cell growth inhibition. Through enzyme detoxification analysis, the toxicological effects of CuNPs are traced to H(2)O(2) generation from CuNPs. Rapid copper release from CuNPs and Cu(I) production are observed. The oxidation of the released Cu(I) has a close relation to H(2)O(2) production, as tris-(hydroxypropyltriazolylmethyl) amine, the specific Cu(I) chelator, can largely protect the cells from the toxicity of CuNPs. In addition, the TEM study shows that CuNPs can be adsorbed and incepted fast by the cells. Comparatively, copper microparticles are relatively stable in the system and practically non-toxic, which indicates the importance of toxic estimation of materials at the nanoscale. In addition, the Cu(II) ion can induce protein damage, membrane damage, and slight DNA damage only at a relatively high concentration. The current study reveals the preliminary mechanism of toxicity of CuNPs, and suggests that the stress-responsive bacterial biosensor array can be used as a simple and promising tool for rapid screening in vitro toxicity of nanoparticles and studying the primary mechanism of the toxicity.
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