Journal article
Beta-subunit-eliminated eHAP expression (BeHAPe) cells reveal subunit regulation of the cardiac voltage-gated sodium channel
The Journal of biological chemistry, Vol.299(9), 105132
09/2023
DOI: 10.1016/j.jbc.2023.105132
PMCID: PMC10506104
PMID: 37544648
Abstract
Voltage-gated sodium (NaV) channels drive the upstroke of the action potential and are comprised of a pore-forming α-subunit and regulatory β-subunits. The β-subunits modulate the gating, trafficking, and pharmacology of the α-subunit. These functions are routinely assessed by ectopic expression in heterologous cells. However, currently available expression systems may not capture the full range of these effects since they contain endogenous β-subunits. To better reveal β-subunit functions, we engineered a human cell line devoid of endogenous NaV β-subunits and their immediate phylogenetic relatives. This new cell line, β-subunit-eliminated eHAP expression cells (BeHAPe), were derived from haploid eHAP cells by engineering inactivating mutations in the β-subunits SCN1B, SCN2B, SCN3B, and SCN4B, and other sub-family members MPZL1, MPZL2, MPZL3, MPZ and JAML. In diploid BeHAPe cells, the cardiac NaV α-subunit, NaV1.5, was highly sensitive to β-subunit modulation and revealed that each β-subunit and even MPZ imparted unique gating properties. Furthermore, combining β1 and β2 with NaV1.5 generated a sodium channel with hybrid properties, distinct from the effects of the individual subunits. Thus, this approach revealed an expanded ability of β-subunits to regulate NaV1.5 activity and can be used to improve the characterization of other α/β NaV complexes.
Details
- Title: Subtitle
- Beta-subunit-eliminated eHAP expression (BeHAPe) cells reveal subunit regulation of the cardiac voltage-gated sodium channel
- Creators
- Annabel Y MinardColin J ClarkChristopher A AhernRobert C Piper
- Resource Type
- Journal article
- Publication Details
- The Journal of biological chemistry, Vol.299(9), 105132
- DOI
- 10.1016/j.jbc.2023.105132
- PMID
- 37544648
- PMCID
- PMC10506104
- NLM abbreviation
- J Biol Chem
- ISSN
- 0021-9258
- eISSN
- 1083-351X
- Grant note
- DOI: 10.13039/100000002, name: National Institutes of Health, award: R01 GM106568, RO1GM058202; DOI: 10.13039/100000041, name: American Diabetes Association; DOI: 10.13039/100001024, name: Roy J. Carver Charitable Trust
- Language
- English
- Electronic publication date
- 08/04/2023
- Date published
- 09/2023
- Academic Unit
- Molecular Physiology and Biophysics; Iowa Neuroscience Institute; Medicine Administration; Internal Medicine
- Record Identifier
- 9984453341402771
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