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PMID: 23129643 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Functional and structural analysis of the human SLO3 pH- and voltage-gated K+ channel.

Leonetti MD, Yuan P, Hsiung Y, Mackinnon R

Abstract

The activation of eukaryotic SLO K(+) channels by intracellular cues, mediated by a cytoplasmic structure called the gating ring, is central to their physiological roles. SLO3 channels are exclusively expressed in mammalian sperm, where variations of intracellular pH are critical to cellular function. Previous studies primarily focused on the mouse SLO3 orthologue and revealed that, in murine sperm, SLO3 mediates a voltage- and alkalization-activated K(+) current essential to male fertility. Here we investigate the activation of the human SLO3 channel by intracellular pH at the functional and structural level. By using electrophysiology in a heterologous system, we show that human SLO3 opens upon intracellular pH increase and that its expression and functional properties are modulated by LRRC52, a testis-specific accessory subunit. We next present the crystal structure of the human SLO3 gating ring. Comparison with the known structures of the corresponding domain from SLO1, a Ca(2+)-activated homologue, suggests that the SLO3 gating ring structure may represent an open state. Together, these results present insights into the function of a protein expected to be critical for human reproduction and provide a framework to study the mechanism of pH gating in SLO3 channels.

MeSH Terms
Animals Humans Hydrogen-Ion Concentration Ion Channel Gating Large-Conductance Calcium-Activated Potassium Channel alpha Subunits/chemistry,metabolism Large-Conductance Calcium-Activated Potassium Channels Membrane Proteins/chemistry,metabolism Mice Models, Molecular Oocytes Potassium Channels, Voltage-Gated/chemistry,metabolism Protein Conformation Protein Subunits/chemistry,metabolism Structural Homology, Protein Xenopus
Chemicals
KCNMA1 protein, human KCNU1 protein, human LRRC52 protein, human Large-Conductance Calcium-Activated Potassium Channel alpha Subunits Large-Conductance Calcium-Activated Potassium Channels Membrane Proteins Potassium Channels, Voltage-Gated Protein Subunits
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Leonetti Manuel D
Laboratory of Molecular Neurobiology and Biophysics and Howard Hughes Medical Institute, The Rockefeller University, New York, NY 10065, USA.
Yuan Peng
Hsiung Yichun
Mackinnon Roderick
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
1091-6490
Published
2012-11-20
Epub
2012-00-05
Pages
19274-9
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC3511096
Subset
IM
Databases
PDB
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