Acta NaturaeActa NaturaeActa Naturae2075-8251Acta Naturae Ltd1082610.32607/20758251-2009-1-1-91-95ArticlesСтатьиResearch ArticleStudying of Membrane Localization of Recombinant Potassium Channels in E.coliStudying of Membrane Localization of Recombinant Potassium Channels in E.coliNekrasovaOgenchem@ibch.ruTagwayAIgnatovaAFeofanovAKirpichnikovMShemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of SciencesBiological Faculty, Lomonosov Moscow State UniversityBiological Faculty, Lomonosov Moscow State UniversityShemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences1506200911NO1 (2009)№1 (2009)919517012020Copyright ©; 2009, Nekrasova O., Tagway A., Ignatova A., Feofanov A., Kirpichnikov M.Copyright ©; 2009, Nekrasova O., Tagway A., Ignatova A., Feofanov A., Kirpichnikov M.2009Nekrasova O., Tagway A., Ignatova A., Feofanov A., Kirpichnikov M.Nekrasova O., Tagway A., Ignatova A., Feofanov A., Kirpichnikov M.https://creativecommons.org/licenses/by/4.0https://actanaturae.ru/2075-8251/article/view/10826

The effective expression of recombinant membrane proteins in E.coli depends upon the targeting and insertion of proteins into the cellular membrane, as well as on those proteins adopting the correct spatial structure. A significant technological problem involves the design of approaches for detecting the location of target proteins within a host cell. Using a hybrid potassium channel KcsA-Kv1.3 as a model, we developed a technological scheme which is suitable for the study of membrane localization in E.coli cells of recombinant proteins containing voltage-gated eukaryotic potassium channels as the functional active site. The scheme involves both biochemical and fluorescent methods for detecting target proteins in the cytoplasmic membrane of E.coli, as well as the study of the ligand-binding activity of membrane-embedded proteins.

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