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Int. J. Mol. Sci. 2018, 19(2), 543; https://doi.org/10.3390/ijms19020543

Determination of Proteinaceous Selenocysteine in Selenized Yeast

Institute of Analytical Sciences, IPREM, UMR 5254, CNRS-UPPA, Hélioparc, 2 Avenue Angot, 64053 Pau, France
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Received: 26 December 2017 / Revised: 29 January 2018 / Accepted: 31 January 2018 / Published: 11 February 2018
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Abstract

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. View Full-Text
Keywords: selenized yeast; selenocysteine; HPLC-ICP MS; HPLC-ESI MS selenized yeast; selenocysteine; HPLC-ICP MS; HPLC-ESI MS
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).
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Bierla, K.; Lobinski, R.; Szpunar, J. Determination of Proteinaceous Selenocysteine in Selenized Yeast. Int. J. Mol. Sci. 2018, 19, 543.

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