Preparation of CuO/ZnO nanocomposite and its application as a cysteine/homocysteine colorimetric and fluorescence detector.

Cysteine and homocysteine play a crucial role in many biological functions but abnormal levels of these amino acids may lead to various forms of pathogenesis. Therefore, selective and easy-to-use methods for the detection of cysteine and homocysteine are essential for the early diagnosis of developing diseases. In this paper we report on a rapid, straightforward and highly selective method for the detection of cysteine (Cys) and homocysteine (Hcy) which uses a CuO/ZnO nanocomposite as a dual colorimetric and fluorometric assay. The presence of Cys and Hcy in a solution of these nanorods (NRs) induces a change in its color from light blue to dark grey which is visible to the naked eye. This is accompanied by a blue shift in the absorption spectra from 725 nm to 650 nm and a decrease in the intensity of CuO/ZnO nanocomposite emission. These changes are ascribed to the reduction of Cu(II) to Cu(0), and the oxidation of cysteine (homocysteine) and subsequent formation of the disulfide bond. This novel assay method does not respond to any other amino-acid which is present in living organisms; therefore the selective determination of cysteine (homocysteine) with a lower analyte limit of 40 µM (4.8 µg mL(-1)) can be carried out in aqueous solutions without the need for any sophisticated instrumentation, fluorophore molecules or complicated procedures.

Cytochrome c conjugated to ZnO-MAA nanoparticles: the study of interaction and influence on protein structure.

Nanoparticle-protein conjugates have potential for numerous applications due to the combination of the properties of both components. In this paper we studied the conjugation of horse heart cytochrome c with ZnO nanoparticles modified by mercaptoacetic acid (MAA) which may be a material with great potential in anticancer therapy as a consequence of synergic effect of both components. Cyt c adsorption to the ZnO-MAA NPs surface was studied by UV-vis spectroscopy and by a dynamic light scattering in various pH. The results indicate that the optimal pH for the association of protein with modified nanoparticles is in range 5.8-8.5 where 90-96% of cytochrome c was assembled on ZnO-MAA nanoparticles. The interaction of proteins with nanoparticles often results in denaturation or loss of protein function. Our observations from UV-vis spectroscopy and circular dichroism performed preserved protein structure after the interaction with modified nanoparticles.

Interaction of cytochrome c with zinc oxide nanoparticles.

The influence of pH on the interaction between horse heart ferricytochrome c (cyt c) and zinc oxide nanoparticles (ZnO NPs) has been studied by a small angle scattering as well as UV-vis and FTIR spectroscopy. The observations showed that the optimal pH for the association of protein with nanoparticles is in pH range 5.0-8.0. Almost no significant change in structure and thermodynamic stability of cytochrome c after the association with 60 nm ZnO NPs was performed by UV-vis and by a circular dichroism spectroscopy.

Alkaline transition of horse heart cytochrome c in the presence of ZnO nanoparticles.

The effect of zinc oxide nanoparticles (ZnO NPs) on cytochrome c (cyt c) in alkaline pH was studied with absorption spectroscopy and UV circular dichroism (CD). Spectral data from UV-vis spectroscopy and circular dichroism indicate only small changes in the native structure of the protein at neutral pH after the interaction with ZnO nanoparticles. The stability around the heme crevice of cyt c and therefore the switch of the axial ligand Met80 to Lys which occurs in conditions of higher pH was proven following the interaction of cytochrome c with ZnO nanoparticles. The formation of cyt c-ZnO NPs complex based on electrostatic attraction was accompanied by a significant increase in the apparent pKa constant of the alkaline transition of cyt c.