engleski

Proteomic responses of Nicotiana tabacum seedlings exposed to differently coated silver nanoparticles

Nanoparticles (NPs) research is currently an area of intense scientific surveys, due to a variety of their potential applications. However, the characteristics making NPs attractive for their exploitation in new products have led to concerns that NPs may pose a risk for environment and human health. Among different available NPs, silver nanoparticles (AgNPs) are of particular interest because of their well-known antibacterial and antifungal properties. They can be found in medical devices, textiles, food packaging, healthcare and household products. Since plants play a significant role in accumulation and biodistribution of many environmentally released substances, they could be influenced by AgNPs, serving as a potential pathway for AgNP-transport and bioaccumulation into food chains. Current proteomics-based techniques offer powerful approaches for the identification of proteins associated with a particular environmental and/or developmental signal, but have been scarcely used on plants to analyse the effects of AgNPs. Our research goal was to determine influence of differently coated AgNPs to tobacco seedlings proteome. Tobacco seedlings were grown from seeds in liquid ½-MS medium supplemented with 100µM citrate-AgNP, PEG-AgNP and BPEI-AgNP. After 10 days proteins from whole seedlings were extracted and loaded for isoelectrofocusing-IEF. IEF was carried out with 700 µg of total protein extract by using non-linear immobilized pH 3-10 gradient strips followed by 2D-PAGE. Proteins were detected with Coomassie brilliant blue. Tryptic peptides were purified using Bravo AsayMAP instumet. 50 protein spots were identified using 4800 Plus MALDI TOF/TOF analyzer. 11 protein spots which were not identified successfully were repeated and subjected to 1-D strong cation-exchange fractionation (SCX) and pH10/pH2.5 C18 2-D fractionation using Bravo AsayMAP instrumet. Until now we successfully purified, fractionated and identified several proteins especially ones with glycine rich domains. With this method we will be able to identify proteins which are difficult to purify and identify with conventional methods.

silver nanoparticles; proteomic response; tobacco seedlings

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