title: Both transmembrane domains of BK β1 subunits are essential to confer the normal phenotype of β1-containing BK channels.
abstract: Voltage/Ca²⁺(i)-gated, large conductance K+ (BK) channels result from tetrameric association of α (slo1) subunits. In most tissues, BK protein complexes include regulatory β subunits that contain two transmembrane domains (TM1, TM2), an extracellular loop, and two short intracellular termini. Four BK β types have been identified, each presenting a rather selective tissue-specific expression profile. Thus, BK β modifies current phenotype to suit physiology in a tissue-specific manner. The smooth muscle-abundant BK β1 drastically increases the channel's apparent Ca²⁺(i) sensitivity. The resulting phenotype is critical for BK channel activity to increase in response to Ca2+ levels reached near the channel during depolarization-induced Ca2+ influx and myocyte contraction. The eventual BK channel activation generates outward K+ currents that drive the membrane potential in the negative direction and eventually counteract depolarization-induced Ca2+ influx. The BK β1 regions responsible for the characteristic phenotype of β1-containing BK channels remain to be identified. We used patch-clamp electrophysiology on channels resulting from the combination of smooth muscle slo1 (cbv1) subunits with smooth muscle-abundant β1, neuron-abundant β4, or chimeras constructed by swapping β1 and β4 regions, and determined the contribution of specific β1 regions to the BK phenotype. At Ca2+ levels found near the channel during myocyte contraction (10 µM), channel complexes that included chimeras having both TMs from β1 and the remaining regions ("background") from β4 showed a phenotype (V(half), τ(act), τ(deact)) identical to that of complexes containing wt β1. This phenotype could not be evoked by complexes that included chimeras combining either β1 TM1 or β1 TM2 with a β4 background. Likewise, β "halves" (each including β1 TM1 or β1 TM2) resulting from interrupting the continuity of the EC loop failed to render the normal phenotype, indicating that physical connection between β1 TMs via the EC loop is also necessary for proper channel function.
authors.affiliation: Department of Pharmacology, College of Medicine, The University of Tennessee Health Science Center, Memphis, Tennessee, United States of America.
authors.forename: Guruprasad
authors.initials: G
authors.lastname: Kuntamallappanavar
authors.affiliation: Departments of Anesthesiology and of Molecular and Medical Pharmacology, University of California Los Angeles, David Geffen School of Medicine, Los Angeles, California, United States of America.
authors.forename: Ligia
authors.initials: L
authors.lastname: Toro
authors.affiliation: Department of Pharmacology, College of Medicine, The University of Tennessee Health Science Center, Memphis, Tennessee, United States of America.
authors.forename: Alex M
authors.initials: AM
authors.lastname: Dopico
mesh_terms: D000818:Animals; D006801:Humans; D051038:Large-Conductance Calcium-Activated Potassium Channel beta Subunits / Q000032:analysis / Q000378:metabolism*; D008564:Membrane Potentials; D032342:Muscle Cells / Q000166:cytology / Q000378:metabolism; D009130:Muscle, Smooth / Q000166:cytology / Q000378:metabolism; D018408:Patch-Clamp Techniques; D017434:Protein Structure, Tertiary; D051381:Rats; D014981:Xenopus
chemical_list: C531905:KCNMB1 protein, human; D051038:Large-Conductance Calcium-Activated Potassium Channel beta Subunits