Your browser doesn't support javascript.
loading
An ancestral MAGUK protein supports the modulation of mammalian voltage-gated Ca2+ channels through a conserved CaVß-like interface.
Segura, Emilie; Mehta, Amrit; Marsolais, Mireille; Quan, Xin R; Zhao, Juan; Sauvé, Rémy; Spafford, J David; Parent, Lucie.
Afiliación
  • Segura E; Département de Pharmacologie et Physiologie, Faculté de Médecine, Canada; Centre de Recherche de l'Institut de Cardiologie de Montréal, Université de Montréal, Montréal, Québec H1T 1C8, Canada.
  • Mehta A; Department of Biology, University of Waterloo, Waterloo, ON, Canada.
  • Marsolais M; Centre de Recherche de l'Institut de Cardiologie de Montréal, Université de Montréal, Montréal, Québec H1T 1C8, Canada.
  • Quan XR; Department of Biology, University of Waterloo, Waterloo, ON, Canada.
  • Zhao J; Centre de Recherche de l'Institut de Cardiologie de Montréal, Université de Montréal, Montréal, Québec H1T 1C8, Canada.
  • Sauvé R; Département de Pharmacologie et Physiologie, Faculté de Médecine, Canada.
  • Spafford JD; Department of Biology, University of Waterloo, Waterloo, ON, Canada.
  • Parent L; Département de Pharmacologie et Physiologie, Faculté de Médecine, Canada; Centre de Recherche de l'Institut de Cardiologie de Montréal, Université de Montréal, Montréal, Québec H1T 1C8, Canada. Electronic address: lucie.parent@umontreal.ca.
Biochim Biophys Acta Biomembr ; 1862(11): 183439, 2020 11 01.
Article en En | MEDLINE | ID: mdl-32814116
Eukaryote voltage-gated Ca2+ channels of the CaV2 channel family are hetero-oligomers formed by the pore-forming CaVα1 protein assembled with auxiliary CaVα2δ and CaVß subunits. CaVß subunits are formed by a Src homology 3 (SH3) domain and a guanylate kinase (GK) domain connected through a HOOK domain. The GK domain binds a conserved cytoplasmic region of the pore-forming CaVα1 subunit referred as the "AID". Herein we explored the phylogenetic and functional relationship between CaV channel subunits in distant eukaryotic organisms by investigating the function of a MAGUK protein (XM_004990081) cloned from the choanoflagellate Salpingoeca rosetta (Sro). This MAGUK protein (Sroß) features SH3 and GK structural domains with a 25% primary sequence identity to mammalian CaVß. Recombinant expression of its cDNA with mammalian high-voltage activated Ca2+ channel CaV2.3 in mammalian HEK cells produced robust voltage-gated inward Ca2+ currents with typical activation and inactivation properties. Like CaVß, Sroß prevents fast degradation of total CaV2.3 proteins in cycloheximide assays. The three-dimensional homology model predicts an interaction between the GK domain of Sroß and the AID motif of the pore-forming CaVα1 protein. Substitution of AID residues Trp (W386A) and Tyr (Y383A) significantly impaired co-immunoprecipitation of CaV2.3 with Sroß and functional upregulation of CaV2.3 currents. Likewise, a 6-residue deletion within the GK domain of Sroß, similar to the locus found in mammalian CaVß, significantly reduced peak current density. Altogether our data demonstrate that an ancestor MAGUK protein reconstitutes the biophysical and molecular features responsible for channel upregulation by mammalian CaVß through a minimally conserved molecular interface.
Asunto(s)
Palabras clave
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas Protozoarias / Canales de Calcio Tipo R / Proteínas de Transporte de Catión / Guanilato-Quinasas Tipo de estudio: Prognostic_studies Límite: Humans Idioma: En Revista: Biochim Biophys Acta Biomembr Año: 2020 Tipo del documento: Article País de afiliación: Canadá Pais de publicación: Países Bajos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas Protozoarias / Canales de Calcio Tipo R / Proteínas de Transporte de Catión / Guanilato-Quinasas Tipo de estudio: Prognostic_studies Límite: Humans Idioma: En Revista: Biochim Biophys Acta Biomembr Año: 2020 Tipo del documento: Article País de afiliación: Canadá Pais de publicación: Países Bajos