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Fluorescent protein-scorpion toxin chimera is a convenient molecular tool for studies of potassium channels

Ion channels play a central role in a host of physiological and pathological processes and are the second largest target for existing drugs. There is an increasing need for reliable tools to detect and visualize particular ion channels, but existing solutions suffer from a number of limitations such...

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Autores principales: Kuzmenkov, Alexey I., Nekrasova, Oksana V., Kudryashova, Kseniya S., Peigneur, Steve, Tytgat, Jan, Stepanov, Alexey V., Kirpichnikov, Mikhail P., Grishin, Eugene V., Feofanov, Alexey V., Vassilevski, Alexander A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5030662/
https://www.ncbi.nlm.nih.gov/pubmed/27650866
http://dx.doi.org/10.1038/srep33314
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author Kuzmenkov, Alexey I.
Nekrasova, Oksana V.
Kudryashova, Kseniya S.
Peigneur, Steve
Tytgat, Jan
Stepanov, Alexey V.
Kirpichnikov, Mikhail P.
Grishin, Eugene V.
Feofanov, Alexey V.
Vassilevski, Alexander A.
author_facet Kuzmenkov, Alexey I.
Nekrasova, Oksana V.
Kudryashova, Kseniya S.
Peigneur, Steve
Tytgat, Jan
Stepanov, Alexey V.
Kirpichnikov, Mikhail P.
Grishin, Eugene V.
Feofanov, Alexey V.
Vassilevski, Alexander A.
author_sort Kuzmenkov, Alexey I.
collection PubMed
description Ion channels play a central role in a host of physiological and pathological processes and are the second largest target for existing drugs. There is an increasing need for reliable tools to detect and visualize particular ion channels, but existing solutions suffer from a number of limitations such as high price, poor specificity, and complicated protocols. As an alternative, we produced recombinant chimeric constructs (FP-Tx) consisting of fluorescent proteins (FP) fused with potassium channel toxins from scorpion venom (Tx). In particular, we used two FP, eGFP and TagRFP, and two Tx, OSK1 and AgTx2, to create eGFP-OSK1 and RFP-AgTx2. We show that these chimeras largely retain the high affinity of natural toxins and display selectivity to particular ion channel subtypes. FP-Tx are displaced by other potassium channel blockers and can be used as an imaging tool in ion channel ligand screening setups. We believe FP-Tx chimeras represent a new efficient molecular tool for neurobiology.
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spelling pubmed-50306622016-09-26 Fluorescent protein-scorpion toxin chimera is a convenient molecular tool for studies of potassium channels Kuzmenkov, Alexey I. Nekrasova, Oksana V. Kudryashova, Kseniya S. Peigneur, Steve Tytgat, Jan Stepanov, Alexey V. Kirpichnikov, Mikhail P. Grishin, Eugene V. Feofanov, Alexey V. Vassilevski, Alexander A. Sci Rep Article Ion channels play a central role in a host of physiological and pathological processes and are the second largest target for existing drugs. There is an increasing need for reliable tools to detect and visualize particular ion channels, but existing solutions suffer from a number of limitations such as high price, poor specificity, and complicated protocols. As an alternative, we produced recombinant chimeric constructs (FP-Tx) consisting of fluorescent proteins (FP) fused with potassium channel toxins from scorpion venom (Tx). In particular, we used two FP, eGFP and TagRFP, and two Tx, OSK1 and AgTx2, to create eGFP-OSK1 and RFP-AgTx2. We show that these chimeras largely retain the high affinity of natural toxins and display selectivity to particular ion channel subtypes. FP-Tx are displaced by other potassium channel blockers and can be used as an imaging tool in ion channel ligand screening setups. We believe FP-Tx chimeras represent a new efficient molecular tool for neurobiology. Nature Publishing Group 2016-09-21 /pmc/articles/PMC5030662/ /pubmed/27650866 http://dx.doi.org/10.1038/srep33314 Text en Copyright © 2016, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Kuzmenkov, Alexey I.
Nekrasova, Oksana V.
Kudryashova, Kseniya S.
Peigneur, Steve
Tytgat, Jan
Stepanov, Alexey V.
Kirpichnikov, Mikhail P.
Grishin, Eugene V.
Feofanov, Alexey V.
Vassilevski, Alexander A.
Fluorescent protein-scorpion toxin chimera is a convenient molecular tool for studies of potassium channels
title Fluorescent protein-scorpion toxin chimera is a convenient molecular tool for studies of potassium channels
title_full Fluorescent protein-scorpion toxin chimera is a convenient molecular tool for studies of potassium channels
title_fullStr Fluorescent protein-scorpion toxin chimera is a convenient molecular tool for studies of potassium channels
title_full_unstemmed Fluorescent protein-scorpion toxin chimera is a convenient molecular tool for studies of potassium channels
title_short Fluorescent protein-scorpion toxin chimera is a convenient molecular tool for studies of potassium channels
title_sort fluorescent protein-scorpion toxin chimera is a convenient molecular tool for studies of potassium channels
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5030662/
https://www.ncbi.nlm.nih.gov/pubmed/27650866
http://dx.doi.org/10.1038/srep33314
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