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Europium-labeled GTP as a general nonradioactive substitute for [35S]GTPγS in high-throughput G protein studies

Europium-labeled GTP as a general nonradioactive substitute for [35S]GTPγS in high-throughput G protein studies

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KOVAL, Alexey, Damir KOPEIN, Vladimir PURVANOV, Vladimir KATANAEV, 2010. Europium-labeled GTP as a general nonradioactive substitute for [35S]GTPγS in high-throughput G protein studies. In: Analytical Biochemistry. 397(2), pp. 202-207. ISSN 0003-2697. eISSN 1096-0309

@article{Koval2010-02-15Europ-14813, title={Europium-labeled GTP as a general nonradioactive substitute for [35S]GTPγS in high-throughput G protein studies}, year={2010}, doi={10.1016/j.ab.2009.10.028}, number={2}, volume={397}, issn={0003-2697}, journal={Analytical Biochemistry}, pages={202--207}, author={Koval, Alexey and Kopein, Damir and Purvanov, Vladimir and Katanaev, Vladimir} }

<rdf:RDF xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#" xmlns:bibo="http://purl.org/ontology/bibo/" xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:dcterms="http://purl.org/dc/terms/" xmlns:xsd="http://www.w3.org/2001/XMLSchema#" > <rdf:Description rdf:about="https://kops.uni-konstanz.de/rdf/resource/123456789/14813"> <dc:contributor>Koval, Alexey</dc:contributor> <dcterms:title>Europium-labeled GTP as a general nonradioactive substitute for [35S]GTPγS in high-throughput G protein studies</dcterms:title> <dc:contributor>Kopein, Damir</dc:contributor> <dc:contributor>Purvanov, Vladimir</dc:contributor> <dcterms:issued>2010-02-15</dcterms:issued> <dc:contributor>Katanaev, Vladimir</dc:contributor> <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-08-18T15:34:41Z</dc:date> <dc:language>eng</dc:language> <bibo:uri rdf:resource="http://kops.uni-konstanz.de/handle/123456789/14813"/> <dc:creator>Koval, Alexey</dc:creator> <dc:creator>Katanaev, Vladimir</dc:creator> <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-08-18T15:34:41Z</dcterms:available> <dc:creator>Kopein, Damir</dc:creator> <dcterms:abstract>[(35)S]GTPgammaS, the nonhydrolyzable radioactive GTP analog, has been a powerful tool in G protein studies and has set the standards in this field of research. However, its radioactive nature imposes clear limitations to its use in regular laboratory practice and in high-throughput experimentation. The europium-labeled GTP analog (Eu-GTP) has been used as an alternative in the analysis of G protein activation by G protein-coupled receptors in cellular membrane preparations. Here we expand the usage of Eu-GTP and show that it can be applied in other types of assays where [(35)S]GTPgammaS has been previously utilized. We demonstrate the applicability of the modified Eu-GTP binding technology to analysis of heterotrimeric and monomeric G proteins of natural and recombinant sources, from different organisms, in assays with soluble proteins and membrane-containing assays of a high-throughput format. The deci-nanomolar K(D) of Eu-GTP for the tested G proteins is similar to that of other fluorescent-modified GTP analogs, while the sensitivity achieved in time-resolved fluorescence analysis of Eu-GTP exceeds that of the radioactive measurements. Overall, the results of our modified Eu-GTP binding assay present Eu-GTP as a general nonradioactive alternative for G protein studies, especially attractive in high-throughput experiments.</dcterms:abstract> <dc:creator>Purvanov, Vladimir</dc:creator> <dc:rights>deposit-license</dc:rights> <dcterms:rights rdf:resource="http://nbn-resolving.org/urn:nbn:de:bsz:352-20140905103605204-4002607-1"/> <dcterms:bibliographicCitation>Publ. in: Analytical Biochemistry ; 397 (2010), 2. - pp. 202-207</dcterms:bibliographicCitation> </rdf:Description> </rdf:RDF>

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