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Detection of Selenoproteins by Laser Ablation Inductively Coupled Plasma Mass Spectrometry (LA-ICP MS) in Immobilized pH Gradient (IPG) Strips

Authors :
Sandra Mounicou
Laurent Chavatte
Jordan Sonet
Institut des sciences analytiques et de physico-chimie pour l'environnement et les materiaux (IPREM)
Université de Pau et des Pays de l'Adour (UPPA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)
CNRS (ATIP program), the Fondation pour la Recherche Médicale (LC), the Ligue Contre le Cancer (Comité de l’Essonne), the programme interdisciplinaire de recherche du CNRS longévité et vieillissement, the Association pour la recherche sur le cancer (grants numbers 4849)
ANR-09-BLAN-0048,SELENOPROTEOME(2009)
Institut des sciences analytiques et de physico-chimie pour l'environnement et les materiaux ( IPREM )
Université de Pau et des Pays de l'Adour ( UPPA ) -Centre National de la Recherche Scientifique ( CNRS )
ANR-09-BLAN-0048,SELENOPROTEOME,Quantitative analysis of the selenoproteome for studies of the kinetics and hierarchy of selenoprotein expression in human cell lines ( 2009 )
Source :
Methods in Molecular Biology, Methods in Molecular Biology, 1661, pp.205-217, 2018, Selenoproteins, ⟨10.1007/978-1-4939-7258-6_15⟩, Selenoproteins, Selenoproteins, 1661, pp.205-217, 2018, 〈10.1007/978-1-4939-7258-6_15〉. 〈https://link.springer.com/protocol/10.1007/978-1-4939-7258-6_15〉, Methods in Molecular Biology ISBN: 9781493972579
Publication Year :
2018
Publisher :
HAL CCSD, 2018.

Abstract

International audience; 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.

Details

Language :
English
ISBN :
978-1-4939-7257-9
ISBNs :
9781493972579
Database :
OpenAIRE
Journal :
Methods in Molecular Biology, Methods in Molecular Biology, 1661, pp.205-217, 2018, Selenoproteins, ⟨10.1007/978-1-4939-7258-6_15⟩, Selenoproteins, Selenoproteins, 1661, pp.205-217, 2018, 〈10.1007/978-1-4939-7258-6_15〉. 〈https://link.springer.com/protocol/10.1007/978-1-4939-7258-6_15〉, Methods in Molecular Biology ISBN: 9781493972579
Accession number :
edsair.doi.dedup.....43057e46305a8443cbba8d7010f7264c
Full Text :
https://doi.org/10.1007/978-1-4939-7258-6_15⟩