Abstract
In contrast to other trace elements that are cofactors of enzymes and removed from proteins under denaturing conditions, Se is covalently bound to proteins when incorporated into selenoproteins, since it is a component of selenocysteine aminoacid. It implies that selenoproteins can undergo several biochemical separation methods in stringent and chaotropic conditions and still maintain the presence of selenium in the primary sequence. This feature has been used to develop a method for the detection of trace levels of human selenoproteins in cell extracts without the use of radioactive isotopes. The selenoproteins are separated as a function of their isoelectric point (pI) using iso-electrofocusing (IEF) electrophoretic strips and detected by laser ablation-inductively coupled plasma mass spectrometry (LA-ICP MS). This method, therefore referred to as IEF-LA-ICP MS, allowed the detection of several selenoproteins in human cell lines, including Gpx1, Gpx4, TXNRD1, TXNRD2, and SELENOF.
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Acknowledgments
This work was supported by the CNRS (ATIP program to LC), the Fondation pour la Recherche Médicale (LC), the Ligue Contre le Cancer (Comité de l’Essonne, LC), the programme interdisciplinaire de recherche du CNRS longévité et vieillissement (LC), the Association pour la recherche sur le cancer [grants numbers 4849, LC] and the Agence Nationale de la Recherche [grant number ANR-09-BLAN-0048 to LC]. JS is a recipient of a Ph.D fellowship from the French Ministry for Research.
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Sonet, J., Mounicou, S., Chavatte, L. (2018). Detection of Selenoproteins by Laser Ablation Inductively Coupled Plasma Mass Spectrometry (LA-ICP MS) in Immobilized pH Gradient (IPG) Strips. In: Chavatte, L. (eds) Selenoproteins. Methods in Molecular Biology, vol 1661. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7258-6_15
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DOI: https://doi.org/10.1007/978-1-4939-7258-6_15
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