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A NOVEL APPROACH FOR THE DETERMINATION OF MEMBRANE PROTEIN EXPRESSION LEVELS BASED ON LC-MS/MS
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SYSNO ASEP 0365269 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title A NOVEL APPROACH FOR THE DETERMINATION OF MEMBRANE PROTEIN EXPRESSION LEVELS BASED ON LC-MS/MS Author(s) Hynek, R. (CZ)
Maršálová, L. (CZ)
Kučková, Š. (CZ)
Koník, P. (CZ)
Šantrůček, J. (CZ)
Martinec, Jan (UEB-Q) RID, ORCID
Novotná, Z. (CZ)
Kodíček, M. (CZ)Source Title Journal of Liquid Chromatography & Related Technologies. - : Taylor & Francis - ISSN 1082-6076
Roč. 34, č. 12 (2011), s. 1004-1013Number of pages 10 s. Language eng - English Country US - United States Keywords expression levels ; isoxaben ; iTRAQ Subject RIV CE - Biochemistry R&D Projects GA203/09/0036 GA ČR - Czech Science Foundation (CSF) LC06034 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) CEZ AV0Z50380511 - UEB-Q (2005-2011) UT WOS 000291807100005 DOI 10.1080/10826076.2011.569808 Annotation Membrane proteins play crucial roles in a number of cellular processes. Thus, the determination of their expression levels under various physiological conditions is crucial in many biological and biochemical studies. However, the extremely hydrophobic nature of proteins possessing transmembrane domains complicates their analysis. Therefore, we developed a new approach toward quantification of membrane proteins in complex mixtures combining reversed-phase chromatography on C4 resin and mass spectrometric techniques using iTRAQ labels. The method was tested on plasma membrane proteins obtained from cell cultures of A.t. by 2-phase partitioning of microsomal fractions. Then, the technique was applied for determination of the influence of the pesticide isoxaben on plasma membrane protein expression levels, particularly plasma membrane Hþ-ATPase. The developed technique seems to be promising for the determination of expression levels of membrane proteins in general. Workplace Institute of Experimental Botany Contact David Klier, knihovna@ueb.cas.cz, Tel.: 220 390 469 Year of Publishing 2012
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