Determination of Proteinaceous Selenocysteine in Selenized Yeast
AbstractA 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
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Bierla, K.; Lobinski, R.; Szpunar, J. Determination of Proteinaceous Selenocysteine in Selenized Yeast. Int. J. Mol. Sci. 2018, 19, 543.
Bierla K, Lobinski R, Szpunar J. Determination of Proteinaceous Selenocysteine in Selenized Yeast. International Journal of Molecular Sciences. 2018; 19(2):543.Chicago/Turabian Style
Bierla, Katarzyna; Lobinski, Ryszard; Szpunar, Joanna. 2018. "Determination of Proteinaceous Selenocysteine in Selenized Yeast." Int. J. Mol. Sci. 19, no. 2: 543.
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