A comparative study of the Se/S substitution in methionine and cysteine in Se-enriched yeast using an inductively coupled plasma mass spectrometry (ICP MS)-assisted proteomics approach

Katarzyna Bierla , Juliusz Bianga , L. Ouerdane , Joanna Szpunar , A. Yiannikouris , Ryszard Łobiński

Abstract

A proteomics approach based on 2D gel electrophoresis followed by HPLC-electrospray Orbitrap MS/MS was developed to investigate the replacement and the degree of the Se/S substitution in methionine and cysteine in Se-rich yeast. Capillary HPLC-inductively coupled plasma mass spectrometry (ICP-MS), employed in parallel to capHPLC–ESI MS, indicated the virtual independence of the ESI MS response of the peptide structure (in the elution range of 30–65% methanol), and hence, the use of ESI MS data to determine the SeCys/Cys and SeMet/Met substitution ratios. For the first time a considerable incorporation of selenocysteine (SeCys) in proteins of the yeast proteome despite the absence of the UGA codon was demonstrated. The SeMet/Met and SeCys/Cys ratios were determined in a large number of peptides (57 and 26, respectively) issued from the tryptic digestion of 19 Se-containing proteins located in the gel by laser ablation-ICP MS imaging. The average Se/S substitution in methionine was 42.9 ± 35.0 and was protein dependent with ratios ranging from 5 to 160 for individual peptides. The substitution of sulphur in cysteine (14.1 ± 4.8%) in the cysteine-containing peptides was relatively similar (ratios from 9 to 23). Taking into account that the cysteine/methionine average ratio (2:1) in the yeast protein fraction, the study allowed the conclusion that 10–15% of selenium present in Se-enriched yeast is in the form of selenocysteine making up the mass balance of selenium species.
Author Katarzyna Bierla - [Universite de Pau et des Pays de L'Adour]
Katarzyna Bierla,,
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, Juliusz Bianga - [Universite de Pau et des Pays de L'Adour]
Juliusz Bianga,,
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, L. Ouerdane - [Universite de Pau et des Pays de L'Adour]
L. Ouerdane,,
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, Joanna Szpunar - [Universite de Pau et des Pays de L'Adour]
Joanna Szpunar,,
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, A. Yiannikouris - [Alltech]
A. Yiannikouris,,
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, Ryszard Łobiński (FC / CAC)
Ryszard Łobiński,,
- Chair Of Analytical Chemistry
Journal seriesJournal of Proteomics, ISSN 1874-3919
Issue year2013
Vol87
Pages26-39
Publication size in sheets0.65
Keywords in EnglishSelenium, Se-substituted methionine, Cysteine, Yeast proteome
ASJC Classification1303 Biochemistry; 1304 Biophysics
DOIDOI:10.1016/j.jprot.2013.05.010
URL http://www.sciencedirect.com/science/article/pii/S1874391913002510
Languageen angielski
File
wdpb_publikacje_pliki_plik_publikacja_1970_org.pdf 1.22 MB
Score (nominal)35
Score sourcejournalList
ScoreMinisterial score = 35.0, 01-02-2020, ArticleFromJournal
Ministerial score (2013-2016) = 35.0, 01-02-2020, ArticleFromJournal
Publication indicators Scopus Citations = 34; WoS Citations = 31; Scopus SNIP (Source Normalised Impact per Paper): 2013 = 1.011; WoS Impact Factor: 2013 = 3.929 (2) - 2013=4.12 (5)
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