Organelle membrane derived patches: reshaping classical methods for new targets
| Autor: | Christian Slomianny, George Shapovalov, Geert Bultynck, Roman Skryma, Anne-Sophie Borowiec, Dmitri V. Gordienko, Gabriel Bidaux, Natalia Prevarskaya, Abigael Ritaine |
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| Přispěvatelé: | Laboratoire de Physiologie Cellulaire : Canaux ioniques, inflammation et cancer - U 1003 (PHYCELL), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille, Cardiovasculaire, métabolisme, diabétologie et nutrition (CarMeN), Hospices Civils de Lyon (HCL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Université de Lyon-Institut National des Sciences Appliquées (INSA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Institut National de la Recherche Agronomique (INRA), Catholic University of Leuven - Katholieke Universiteit Leuven (KU Leuven), Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Hospices Civils de Lyon (HCL)-Institut National de la Santé et de la Recherche Médicale (INSERM), bidaux, gabriel |
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Patch-Clamp Techniques [SDV]Life Sciences [q-bio] lcsh:Medicine [SDV.BC]Life Sciences [q-bio]/Cellular Biology Cell Fractionation Article 03 medical and health sciences Cell Line Tumor Organelle Humans Inositol 1 4 5-Trisphosphate Receptors Patch clamp lcsh:Science [SDV.BC] Life Sciences [q-bio]/Cellular Biology Ion channel Organelles Multidisciplinary Chemistry HEK 293 cells lcsh:R Intracellular Membranes Cell biology Organelle membrane Electrophysiology 030104 developmental biology Membrane HEK293 Cells Proto-Oncogene Proteins c-bcl-2 lcsh:Q Intracellular |
| Zdroj: | Scientific Reports Scientific Reports, Nature Publishing Group, 2017, 7, pp.14082. ⟨10.1038/s41598-017-13968-y⟩ Scientific Reports, Nature Publishing Group, 2017, 7 (1), pp.14082. ⟨10.1038/s41598-017-13968-y⟩ Scientific Reports, Vol 7, Iss 1, Pp 1-14 (2017) Scientific Reports, 2017, 7, pp.14082. ⟨10.1038/s41598-017-13968-y⟩ |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/s41598-017-13968-y⟩ |
| Popis: | Intracellular ion channels are involved in multiple signaling processes, including such crucial ones as regulation of cellular motility and fate. With 95% of the cellular membrane belonging to intracellular organelles, it is hard to overestimate the importance of intracellular ion channels. Multiple studies have been performed on these channels over the years, however, a unified approach allowing not only to characterize their activity but also to study their regulation by partner proteins, analogous to the patch clamp “golden standard”, is lacking. Here, we present a universal approach that combines the extraction of intracellular membrane fractions with the preparation of patchable substrates that allows to characterize these channels in endogenous protein environment and to study their regulation by partner proteins. We validate this method by characterizing activity of multiple intracellular ion channels localized to different organelles and by providing detailed electrophysiological characterization of the regulation of IP3R activity by endogenous Bcl-2. Thus, after synthesis and reshaping of the well-established approaches, organelle membrane derived patch clamp provides the means to assess ion channels from arbitrary cellular membranes at the single channel level. |
| Databáze: | OpenAIRE |
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