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Determination of Proteinaceous Selenocysteine in Selenized Yeast

A method for the quantitation of proteinaceous selenocysteine (SeCys) in Se-rich yeast was developed. The method is based on the reduction of the Se-Se and S-Se bridges with dithiotretiol, derivatization with iodoacetamide (carbamidomethylation), followed by HPLC-ICP MS. The chromatographic conditio...

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Autores principales: Bierla, Katarzyna, Lobinski, Ryszard, Szpunar, Joanna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5855765/
https://www.ncbi.nlm.nih.gov/pubmed/29439473
http://dx.doi.org/10.3390/ijms19020543
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author Bierla, Katarzyna
Lobinski, Ryszard
Szpunar, Joanna
author_facet Bierla, Katarzyna
Lobinski, Ryszard
Szpunar, Joanna
author_sort Bierla, Katarzyna
collection PubMed
description A method for the quantitation of proteinaceous selenocysteine (SeCys) in Se-rich yeast was developed. The method is based on the reduction of the Se-Se and S-Se bridges with dithiotretiol, derivatization with iodoacetamide (carbamidomethylation), followed by HPLC-ICP MS. The chromatographic conditions were optimized for the total recovery of the proteinaceous selenocysteine, the minimum number of peaks in the chromatogram (reduction of derivatization products of other Se-species present) and the baseline separation. A typical chromatogram of a proteolytic digest of selenized yeast protein consisted of up to five peaks (including SeMet, carbamidomethylated (CAM)-SeCys, and Se(CAM)(2)) identified by retention time matching with available standards and electrospray MS. Inorganic selenium non-specifically attached to proteins and selenomethionine could be quantified (in the form of Se(CAM)(2)) along with SeCys. Selenocysteine, selenomethionine, inorganic selenium, and the water soluble-metabolite fraction accounted for the totality of selenium species in Se-rich yeast.
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spelling pubmed-58557652018-03-20 Determination of Proteinaceous Selenocysteine in Selenized Yeast Bierla, Katarzyna Lobinski, Ryszard Szpunar, Joanna Int J Mol Sci Article A method for the quantitation of proteinaceous selenocysteine (SeCys) in Se-rich yeast was developed. The method is based on the reduction of the Se-Se and S-Se bridges with dithiotretiol, derivatization with iodoacetamide (carbamidomethylation), followed by HPLC-ICP MS. The chromatographic conditions were optimized for the total recovery of the proteinaceous selenocysteine, the minimum number of peaks in the chromatogram (reduction of derivatization products of other Se-species present) and the baseline separation. A typical chromatogram of a proteolytic digest of selenized yeast protein consisted of up to five peaks (including SeMet, carbamidomethylated (CAM)-SeCys, and Se(CAM)(2)) identified by retention time matching with available standards and electrospray MS. Inorganic selenium non-specifically attached to proteins and selenomethionine could be quantified (in the form of Se(CAM)(2)) along with SeCys. Selenocysteine, selenomethionine, inorganic selenium, and the water soluble-metabolite fraction accounted for the totality of selenium species in Se-rich yeast. MDPI 2018-02-11 /pmc/articles/PMC5855765/ /pubmed/29439473 http://dx.doi.org/10.3390/ijms19020543 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Bierla, Katarzyna
Lobinski, Ryszard
Szpunar, Joanna
Determination of Proteinaceous Selenocysteine in Selenized Yeast
title Determination of Proteinaceous Selenocysteine in Selenized Yeast
title_full Determination of Proteinaceous Selenocysteine in Selenized Yeast
title_fullStr Determination of Proteinaceous Selenocysteine in Selenized Yeast
title_full_unstemmed Determination of Proteinaceous Selenocysteine in Selenized Yeast
title_short Determination of Proteinaceous Selenocysteine in Selenized Yeast
title_sort determination of proteinaceous selenocysteine in selenized yeast
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5855765/
https://www.ncbi.nlm.nih.gov/pubmed/29439473
http://dx.doi.org/10.3390/ijms19020543
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