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Functional analyses of heteromeric human PIEZO1 Channels
Radhakrishnan Gnanasambandam1, Chilman Bae1, Lynn Ziegler1
1Department of Physiology and Biophysics, State University of New York at Buffalo, Buffalo, United States of America.
Plos One
|November 22, 2018
Summary
Mechanosensitive channels PIEZO1 and PIEZO2 form functional heteromers. Co-expressed PIEZO1 subunits and PIEZO1/PIEZO2 combinations yield distinct conductances, suggesting domain segregation rather than random mixing.
Area of Science:
- Cellular Biophysics
- Ion Channel Physiology
Background:
- PIEZO1 and PIEZO2 are essential mechanosensitive channels (MSCs) involved in cellular mechanotransduction.
- Mutations in PIEZO channels are linked to various human diseases, highlighting their critical physiological roles.
Purpose of the Study:
- To investigate the formation and properties of functional heteromeric channels involving PIEZO1 subunits.
- To understand how different PIEZO1 mutants assemble into functional complexes and influence channel activity.
Main Methods:
- Utilized site-directed mutagenesis to create PIEZO1 mutants (E2117K, E2117D, E2117A).
- Performed whole-cell electrophysiology to record currents from homomeric and heteromeric channel constructs.
- Employed high-resolution optical imaging of fluorescently-tagged PIEZO channels.
Main Results:
- PIEZO1 homomers with mutations E2117K, E2117A, and E2117D exhibited distinct conductances and inactivation kinetics.
- Co-expression of PIEZO1 mutants generated novel heteromeric channel species with unique electrophysiological properties.
- Electrophysiological data from PIEZO1 heteromers and PIEZO1/PIEZO2 co-expression fit a linear combination model of homomeric currents.
- Optical imaging revealed segregation of coexpressed PIEZO1 subunits into discrete domains.
Conclusions:
- Functional PIEZO1 heteromers can form with distinct conductances, differing from their respective homomers.
- The assembly of functional PIEZO1 and PIEZO1/PIEZO2 channels likely involves non-random organization and domain segregation of subunits.
- Findings suggest that heteromer formation is not a result of freely diffusing, randomly mixed monomers in vitro.
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