Aloria, Kerman

Arizmendi, Jesús M.

Azkargorta, Mikel

Fullaondo, Asier

Zubiaga, J.M.

Repositorio Institucional de la Universidad de Córdoba

S6. Animal, Plant and Microbial Proteomics Posters192P. 119DIFFERENTIAL PROTEIN EXPRESSION ANALYSIS OF ACTIVATED E2F2-/- T LYMPHOCYTES BY MEANS OF A LABEL-FREE QUANTIFICATION METHODM. Azkargorta1, K. Aloria2, A.M. zubiaga3, J.M. Arizmendi1, A. Fullaondo31Department of Biochemistry and Molecular Biology, University of the Basque Country, Leioa, Bizkaia; 2Proteomics Unit, SGIker, University of the Basque Country, Leioa, Bizkaia; 3Department of Genetics, Physical Anthropology and Animal Physiology, University of the Basque Country, Leioa, BizkaiaDifferential proteomics has traditionally relied on two-dimensional electrophoresis (2DE). 2DE is a powerful technique that allows the separation and analysis of hundreds of proteins in a single gel, leading to the analysis of global expression patterns. 2DE, however, has a number of inherent limitations and disadvantages. To overcome these limitations several MS-based gel-free techniques have been developed. Among these, a label-free absolute quantification method based on the three most intense peptides at each protein developed by Waters (Hi3) is very simple and affordable.E2F family of transcription factors (E2F1-8) are crucial regulators of cell-cycle and proliferation. Characterization of mice lacking functional E2F2 transcription factor (E2F2-/-) showed the development of an autoimmune syndrome due to the presence of hypersensible and hyperreactive peripheral T lymphocytes. These T lymphocytes hyperproliferated upon antigenic stimulation, and showed an accelerated entry to the S phase of cell cycle, suggesting a role for E2F2 in the repression of G1/S transition. Previous effort in our lab has allowed us the characterization of E2F2-lacking T lymphocyte’s expression patterns on a 2DE-based approach.In this work we characterize E2F2-/- activated T lymphocyte’s protein expression patterns by Hi3 absolute quantification method. Hi3 analysis of these extracts revealed the deregulation of some interesting cell-cycle and E2F-related proteins, proving the usefulness of this technique for differential expression analysis. Moreover, results obtained by 2DE and Hi3 analysis of activated E2F2-/- T lymphocytes are compared, suggesting that both approaches can provide complementary viewpoints in differential proteomics analyses.

Differential protein expression analysis of activated E2F2-/-T lymphocytes by means of label-free quantification method

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Repositorio Institucional de la Universidad de Córdoba