Direct interaction of CaVß with actin up-regulates L-type calcium currents in HL-1 cardiomyocytes.
J Biol Chem
; 290(8): 4561-4572, 2015 Feb 20.
Article
em En
| MEDLINE
| ID: mdl-25533460
ABSTRACT
Expression of the ß-subunit (CaVß) is required for normal function of cardiac L-type calcium channels, and its up-regulation is associated with heart failure. CaVß binds to the α1 pore-forming subunit of L-type channels and augments calcium current density by facilitating channel opening and increasing the number of channels in the plasma membrane, by a poorly understood mechanism. Actin, a key component of the intracellular trafficking machinery, interacts with Src homology 3 domains in different proteins. Although CaVß encompasses a highly conserved Src homology 3 domain, association with actin has not yet been explored. Here, using co-sedimentation assays and FRET experiments, we uncover a direct interaction between CaVß and actin filaments. Consistently, single-molecule localization analysis reveals streaklike structures composed by CaVß2 that distribute over several micrometers along actin filaments in HL-1 cardiomyocytes. Overexpression of CaVß2-N3 in HL-1 cells induces an increase in L-type current without altering voltage-dependent activation, thus reflecting an increased number of channels in the plasma membrane. CaVß mediated L-type up-regulation, and CaVß-actin association is prevented by disruption of the actin cytoskeleton with cytochalasin D. Our study reveals for the first time an interacting partner of CaVß that is directly involved in vesicular trafficking. We propose a model in which CaVß promotes anterograde trafficking of the L-type channels by anchoring them to actin filaments in their itinerary to the plasma membrane.
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Texto completo:
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Regulação para Cima
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Actinas
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Sinalização do Cálcio
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Canais de Cálcio Tipo L
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Miócitos Cardíacos
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Modelos Biológicos
Limite:
Animals
Idioma:
En
Revista:
J Biol Chem
Ano de publicação:
2015
Tipo de documento:
Article