TY - JOUR
T1 - β1-subunit-induced structural rearrangements of the Ca2+- and voltage-activated K+ (BK) channel
AU - Castillo, Juan P.
AU - Sánchez-Rodríguez, Jorge E.
AU - Hyde, H. Clark
AU - Zaelzer, Cristian A.
AU - Aguayo, Daniel
AU - Sepúlveda, Romina V.
AU - Luk, Louis Y P
AU - Kent, Stephen B H
AU - Gonzalez-Nilo, Fernando D.
AU - Bezanilla, Francisco
AU - Latorre, Ramón
N1 - Funding Information:
We thank Bobo Dang (University of Chicago) for technical assistance with peptide purification and characterization. This research was supported by Fondo Nacional de Desarrollo Científico y Tecnológico Grants 1110430 and 1150273 (to R.L.), 1131003 (to F.D.G.-N.), and 11130576 (to D.A.); Anillo Grant ACT-1107, Comisión Nacional de Investigación Científica y Tecnológica (CONICYT) - Programa de Investigación Asociativa (PIA) (to F.D.G.-N.); NIH Grant GM030376 (to F.B.); and NIH U54GM087519 (to F.B.); a CONACYT postdoctoral fellowship, Mexican Government (to J.E.S.-R.); CONICYT Graduate Fellowship 21090197 and Grant AT-24121240 (to J.P.C.); and CONICYT Grant 21130631, Chilean Government (to R.S.). The Centro Interdisciplinario de Neurociencia de Valparaiso is a Millennium Institute supported by the Millennium Scientific Initiative of the Chilean Ministry of Economy, Development, and Tourism (P029-022-F).
PY - 2016/6/7
Y1 - 2016/6/7
N2 - Large-conductance Ca2+- 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.
AB - Large-conductance Ca2+- 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.
KW - BK channels
KW - Lanthanide resonance energy transfer
KW - β 1-subunit
UR - http://www.scopus.com/inward/record.url?scp=84973316259&partnerID=8YFLogxK
U2 - 10.1073/pnas.1606381113
DO - 10.1073/pnas.1606381113
M3 - Article
AN - SCOPUS:84973316259
SN - 0027-8424
VL - 113
SP - E3231-E3239
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 23
ER -