2. Academic Validation
  3. β1-subunit-induced structural rearrangements of the Ca2+- and voltage-activated K+ (BK) channel

β1-subunit-induced structural rearrangements of the Ca2+- and voltage-activated K+ (BK) channel

  • Proc Natl Acad Sci U S A. 2016 Jun 7;113(23):E3231-9. doi: 10.1073/pnas.1606381113.
Juan P Castillo 1,  Jorge E Sánchez-Rodríguez 2,  H Clark Hyde 3,  Cristian A Zaelzer 4,  Daniel Aguayo 5,  Romina V Sepúlveda 6,  Louis Y P Luk 7,  Stephen B H Kent 8,  Fernando D Gonzalez-Nilo 5,  Francisco Bezanilla 9,  Ramón Latorre 10
Affiliations

Affiliations

  • 1 Centro Interdisciplinario de Neurociencias de Valparaíso, Facultad de Ciencias, Universidad de Valparaíso, Valparaiso 2360103, Chile; Departamento de Biologia, Facultad de Ciencias, Universidad de Chile, Santiago 7800003, Chile;
  • 2 Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL 60637; Departamento de Física, Universidad de Guadalajara, Guadalajara, Jalisco, 44430, Mexico;
  • 3 Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL 60637;
  • 4 Centro Interdisciplinario de Neurociencias de Valparaíso, Facultad de Ciencias, Universidad de Valparaíso, Valparaiso 2360103, Chile;
  • 5 Centro Interdisciplinario de Neurociencias de Valparaíso, Facultad de Ciencias, Universidad de Valparaíso, Valparaiso 2360103, Chile; Universidad Andrés Bello, Center for Bioinformatics and Integrative Biology, Facultad de Ciencias Biológicas, Santiago 8370146, Chile;
  • 6 Universidad Andrés Bello, Center for Bioinformatics and Integrative Biology, Facultad de Ciencias Biológicas, Santiago 8370146, Chile;
  • 7 Department of Chemistry, University of Chicago, Chicago, IL 60637.
  • 8 Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL 60637; Department of Chemistry, University of Chicago, Chicago, IL 60637.
  • 9 Centro Interdisciplinario de Neurociencias de Valparaíso, Facultad de Ciencias, Universidad de Valparaíso, Valparaiso 2360103, Chile; Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL 60637; [email protected] [email protected].
  • 10 Centro Interdisciplinario de Neurociencias de Valparaíso, Facultad de Ciencias, Universidad de Valparaíso, Valparaiso 2360103, Chile; [email protected] [email protected].
PMID: 27217576 DOI: 10.1073/pnas.1606381113
Abstract

Large-conductance Ca(2+)- and voltage-activated K(+) (BK) channels are involved in a large variety of physiological processes. Regulatory β-subunits are one of the mechanisms responsible for creating BK channel diversity fundamental to the adequate function of many tissues. However, little is known about the structure of its voltage sensor domain. Here, we present the external architectural details of BK channels using lanthanide-based resonance energy transfer (LRET). We used a genetically encoded lanthanide-binding tag (LBT) to bind terbium as a LRET donor and a fluorophore-labeled iberiotoxin as the LRET acceptor for measurements of distances within the BK channel structure in a living cell. By introducing LBTs in the extracellular region of the α- or β1-subunit, we determined (i) a basic extracellular map of the BK channel, (ii) β1-subunit-induced rearrangements of the voltage sensor in α-subunits, and (iii) the relative position of the β1-subunit within the α/β1-subunit complex.

Keywords

BK channels; lanthanide resonance energy transfer; β1-subunit.

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