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Issue 6, 2015
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Identification and quantification of seleno-proteins by 2-DE-SR-XRF in selenium-enriched yeasts

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Abstract

A comprehensive approach that can identify and quantify selenium (Se) in seleno-proteins in Se-enriched yeast was developed. The Se-containing compounds in Se-enriched yeast were first extracted and then the fraction of Se-containing proteins in the supernatant was analyzed by 2-dimensional electrophoresis (2-DE) and synchrotron radiation X-ray fluorescence (SR-XRF). The detection limit (DL) of SR-XRF analysis for Se quantification in Se-containing proteins after 2-DE separation was calculated to be 0.20 µg g−1, which is suitable for Se quantification in the Se-containing spots present on the 2-D gel. After being scanned by SR-XRF, only spots with a mean Se content exceeding twice the DL of SR-XRF were considered to be seleno-proteins. In this way, a total of 157 Se-containing spots in the gel were visually distinguished. Se contents in all the Se-containing proteins of different molecular weights were quantified. The total Se content on the 2-D gel was calculated to be 126.56 µg g−1, which covered most of the seleno-proteins on the 2-D gel.

Graphical abstract: Identification and quantification of seleno-proteins by 2-DE-SR-XRF in selenium-enriched yeasts

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Publication details

The article was received on 09 Feb 2015, accepted on 23 Mar 2015 and first published on 23 Mar 2015


Article type: Technical Note
DOI: 10.1039/C5JA00043B
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J. Anal. At. Spectrom., 2015,30, 1408-1413

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    Identification and quantification of seleno-proteins by 2-DE-SR-XRF in selenium-enriched yeasts

    J. Zhao, Y. Pu, Y. Gao, X. Peng, Y. Li, X. Xu, B. Li, N. Zhu, J. Dong, G. Wu and Y. Li, J. Anal. At. Spectrom., 2015, 30, 1408
    DOI: 10.1039/C5JA00043B

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