Servizio Comunitario di Informazione in materia di Ricerca e Sviluppo - CORDIS

Final Activity Report Summary - PROFACT (Protein factory for kinase substrate searching and protein detecting microarray development)

In biological terms, the 21st century is the era of postgenomics research facing the challenging task of untangling and revealing the function of subtle protein networks of living organisms. The large scale studies urge invention of high-throughput approaches capable coping with analysis of thousands of proteins. This new field of biology -the proteomics- largely depends on availability of ample amount of a large number of proteins. According to the objectives of the proposal, we set up a wheat germ extract based cell free protein translation system that meets the requirements of proteomics by producing proteins of interest rapidly and efficiently.

The established protein translation system was applied to produce active protein kinases for identification of their putative substrates. We aim at isolation of target molecules of protein kinases located in mitochondria, since the mitochondria is central cell organ not just as main energy converter but also as central signal integrator. Revealing protein phosphorylation pathways in mitochondria will contribute to understanding of regulation of programmed cell death, an important procedure in development and eradicating tumour cells. In order to implement this task plant mitochondrial protein extracts have been purified and the produced protein kinase will be added to these extracts to label the target molecules.

An equally important objective of the proposal is the isolation of aptamers against proteins. Aptamers are short oligonucleotides binding the target proteins and macromolecules with high specificity and affinity, thus, rivals of antibodies in many applications like, protein and macromolecule detection, affinity purification, immuno-affinity labelling and even used as human therapeutics. In our laboratory, a random aptamer library was designed and used for identification of specific oligonucleotides for protein kinases and mycotoxins. In both cases, these works resulted in isolation of aptamers binding to the target molecules, and possible applications of these oligonucleotides are currently tested.

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Muegyetem rkp. 3.
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