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Identification of selenium-containing proteins in selenium-rich yeast aqueous extract by 2D gel electrophoresis, nanoHPLC-ICP MS and nanoHPLC-ESI MS/MS

Tastet, Laure and Schaumloeffel, Dirk and Bouyssiere, Brice and Lobinski, Ryszard (2008) Identification of selenium-containing proteins in selenium-rich yeast aqueous extract by 2D gel electrophoresis, nanoHPLC-ICP MS and nanoHPLC-ESI MS/MS. TALANTA, 75 (4). pp. 1140-1145.

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Abstract

An approach based on the consecutive use of nanoHPLC-ICP collision cell MS and nanoHPLC-electrospray MS was proposed for the analysis of water-soluble selenium-containing proteins in selenium-rich yeast after their separation by 2D gel electrophoresis (GE). An ultrasonic probe was employed for fast protein extraction avoiding sample heating and thus reducing the risk of protein degradation. The efficiency of different extraction steps were critically evaluated by total selenium analysis and size-exclusion chromatography (SEC)-ICP MS. Prior to electrophoresis proteins were purified by acetone precipitation. The protein-containing spots from 2D GE were excised and digested with trypsin. The digests obtained were analyzed by nanoHPLC-ICP MS in order to check for the presence of selenium-containing peptides; this allowed the detection of target proteins for further analyses (two out of five spots). The subsequent analyses of the selected digests by nanoHPLC-ES MS/MS allowed the attribution of amino acid sequences to peaks detected by ICP MS revealing the presence of two selenium-containing proteins: SIP 18 and HSP 12. (c) 2008 Elsevier B.V. All rights reserved.

Item Type:Article
Uncontrolled Keywords:2D gel electrophoresis; NanoHPLC; ICP MS; electrospray MS/MS; selenium-containing proteins; selenium-rich yeast
Subjects:Material Science > Nanofabrication processes and tools
Biomedical Science > Nanobiotechnology
ID Code:746
Deposited By:SPI
Deposited On:26 Jan 2009 15:11
Last Modified:26 Jan 2009 15:11

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