Abstract
Selenium (Se) is an essential component of genetically encoded selenoproteins, in the form of a rare amino acid, namely the selenocysteine (Sec). Radioactive 75Se has been widely used to trace selenoproteins in vitro and in vivo (cell models and animals). Alternatively, its unique isotopic pattern can be used to detect and characterize nonradioactive Se-compounds in cellular extracts using molecular or elemental mass spectrometry at ppm levels. However, when studying trace levels of Se-compounds, such as selenoproteins (ppt levels), the distribution of the signal between its six naturally abundant isotopes reduces its sensitivity. Here, we describe the use of isotopically enriched forms of Se as an alternative strategy to radioactive 75Se, for the labeling and tracing of selenoproteins in cultured cell lines.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Kryukov GV, Castellano S, Novoselov SV, Lobanov AV, Zehtab O, Guigo R, Gladyshev VN (2003) Characterization of mammalian selenoproteomes. Science 300(5624):1439–1443
Touat-Hamici Z, Legrain Y, Sonet J, Bulteau A-L, Chavatte L (2016) Alteration of selenoprotein expression during stress and in aging. In: Hatfield DL, Su D, Tsuji PA, Gladyshev VN (eds) Selenium: its molecular biology and role in human health, 4th edn. Springer Science+Business Media, LLC, New York, pp 539–551
Sonet J, Bulteau A-L, Chavatte L (2016) Selenium and selenoproteins in human health and diseases. In: Michalke B (ed) Metallomics: analytical techniques and speciation methods. Wiley-VCH Verlag GmbH & Co KGaA, Weinheim, Germany, pp 364–381. doi:10.1002/9783527694907.ch13
Labunskyy VM, Hatfield DL, Gladyshev VN (2014) Selenoproteins: molecular pathways and physiological roles. Physiol Rev 94(3):739–777. doi: 94/3/739 [pii] 10.1152/physrev.00039.2013
Latrèche L, Chavatte L (2008) Selenium incorporation into selenoproteins, implications in human health. Met Ions Biol Med X:731–737
Papp LV, Lu J, Holmgren A, Khanna KK (2007) From selenium to selenoproteins: synthesis, identity, and their role in human health. Antioxid Redox Signal 9(7):775–806
Bulteau A-L, Chavatte L (2015) Update on selenoprotein biosynthesis. Antioxid Redox Signal 23(10):775–794
Latreche L, Duhieu S, Touat-Hamici Z, Jean-Jean O, Chavatte L (2012) The differential expression of glutathione peroxidase 1 and 4 depends on the nature of the SECIS element. RNA Biol 9(5):681–690
Latreche L, Jean-Jean O, Driscoll DM, Chavatte L (2009) Novel structural determinants in human SECIS elements modulate the translational recoding of UGA as selenocysteine. Nucleic Acids Res 37(17):5868–5880
Driscoll DM, Copeland PR (2003) Mechanism and regulation of selenoprotein synthesis. Annu Rev Nutr 23:17–40
Touat-Hamici Z, Legrain Y, Bulteau AL, Chavatte L (2014) Selective up-regulation of human selenoproteins in response to oxidative stress. J Biol Chem 289(21):14750–14761. doi: M114.551994 [pii] 10.1074/jbc.M114.551994
Legrain Y, Touat-Hamici Z, Chavatte L (2014) Interplay between selenium levels, selenoprotein expression, and replicative senescence in WI-38 human fibroblasts. J Biol Chem 289(9):6299–6310
Gladyshev VN, Arner ES, Berry MJ, Brigelius-Flohe R, Bruford EA, Burk RF, Carlson BA, Castellano S, Chavatte L, Conrad M, Copeland PR, Diamond AM, Driscoll DM, Ferreiro A, Flohe L, Green FR, Guigo R, Handy DE, Hatfield DL, Hesketh J, Hoffmann PR, Holmgren A, Hondal RJ, Howard MT, Huang K, Kim H-Y, Kim IY, Kohrle J, Krol A, Kryukov GV, Lee BJ, Lee BC, Lei XG, Liu Q, Lescure A, Lobanov AV, Loscalzo J, Maiorino M, Mariotti M, Sandeep Prabhu K, Rayman MP, Rozovsky S, Salinas G, Schmidt EE, Schomburg L, Schweizer U, Simonovic M, Sunde RA, Tsuji PA, Tweedie S, Ursini F, Whanger PD, Zhang Y (2016) Selenoprotein gene nomenclature. J Biol Chem 291(46):24036–24040
Bianga J, Touat-Hamici Z, Bierla K, Mounicou S, Szpunar J, Chavatte L, Lobinski R (2014) Speciation analysis for trace levels of selenoproteins in cultured human cells. J Proteome 108:316–324. doi:10.1016/j.jprot.2014.05.025
Mounicou S, Bierla K, Szpunar J Detection and determination of heteroatom-containing molecules by HPLC: inductively coupled plasma mass spectrometry. In: Shalliker RA (ed) Hyphenated and alternative methods of detection in chromatography, vol 2011. CRC Press, Boca Raton, FL, pp 161–185
Mounicou S, Szpunar J, Lobinski R (2009) Metallomics: the concept and methodology. Chem Soc Rev 38(4):1119–1138. doi:10.1039/b713633c
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.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Sonet, J., Mounicou, S., Chavatte, L. (2018). Nonradioactive Isotopic Labeling and Tracing of Selenoproteins in Cultured Cell Lines. 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_14
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7258-6_14
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7257-9
Online ISBN: 978-1-4939-7258-6
eBook Packages: Springer Protocols