Quantitative proteomic analysis of lung carcinoma cell line transfected by mutant p53 gene sequences

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Authors	BOUCHAL Pavel HRSTKA Roman HERNYCHOVÁ Lenka LINK Marek TAMBOR Vojtěch HOLEŠÍNSKÁ Pavlína VOJTĚŠEK Bořivoj
Year of publication	2009
Type	Conference abstract
MU Faculty or unit	Faculty of Science
Citation
Description	The present study constitutes the use of a quantitative LC-MS/MS based proteomic approach for the comparative proteomic analysis of lung carcinoma cell line H1299 steady transfected by two different mutant p53 gene sequences (hot-spot mutations at codon positions 175 and 273, respectively). The proteomics analysis method involved iTRAQ 8-plex stable isotope labeling of tryptic peptides, their separation via off-line coupled two-dimensional liquid chromatography (SCX and RP-LC) followed by tandem mass spectrometry on the MALDI-TOF/TOF platform (iTRAQ-2DLC-MS/MS). The principal aims of this study were: (1) to define the protein spectrum detectable using this approach, and (2) to quantitative compare the proteomic profiles of cells, which differ only in type of p53 mutation after 17-(allylamino)-17-demethoxygeldanamycin (17-AAG) treatment. The study resulted in the reproducible identification of 411 non-redundant proteins.
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