Abstract：
: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.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Shapovalov G,Ritaine A,Bidaux G,Slomianny C,Borowiec AS,Gordienko D,Bultynck G,Skryma R,Prevarskaya Ndoi
10.1038/s41598-017-13968-ysubject
Has Abstractpub_date
2017-10-26 00:00:00pages
14082issue
1issn
2045-2322pii
10.1038/s41598-017-13968-yjournal_volume
7pub_type
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