A NOVEL APPROACH FOR THE DETERMINATION OF MEMBRANE PROTEIN EXPRESSION LEVELS BASED ON LC-MS/MS

0365269 - ÚEB 2012 RIV US eng J - Journal Article
Hynek, R. - Maršálová, L. - Kučková, Š. - Koník, P. - Šantrůček, J. - Martinec, Jan - Novotná, Z. - Kodíček, M.
A NOVEL APPROACH FOR THE DETERMINATION OF MEMBRANE PROTEIN EXPRESSION LEVELS BASED ON LC-MS/MS.
Journal of Liquid Chromatography & Related Technologies. Roč. 34, č. 12 (2011), s. 1004-1013. ISSN 1082-6076. E-ISSN 1520-572X
R&D Projects: GA ČR GA203/09/0036; GA MŠMT(CZ) LC06034
Institutional research plan: CEZ:AV0Z50380511
Keywords : expression levels * isoxaben * iTRAQ
Subject RIV: CE - Biochemistry
Impact factor: 0.706, year: 2011

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.
Permanent Link: http://hdl.handle.net/11104/0200552