Structure and electromechanical coupling of a voltage-gated Na⁺/H⁺ exchanger

Voltage-sensing domains control the activation of voltage-gated ion channels, with a few exceptions¹. One such exception is the sperm-specific Na⁺/H⁺ exchanger SLC9C1, which is the only known transporter to be regulated by voltage-sensing domains^2-5. After hyperpolarization of sperm flagella, SLC9C1 becomes active, causing pH alkalinization and CatSper Ca²⁺ channel activation, which drives chemotaxis^2,6. SLC9C1 activation is further regulated by cAMP^2,7, which is produced by soluble adenyl cyclase (sAC). SLC9C1 is therefore an essential component of the pH-sAC-cAMP signalling pathway in metazoa^8,9, required for sperm motility and fertilization⁴. Despite its importance, the molecular basis of SLC9C1 voltage activation is unclear. Here we report cryo-electron microscopy (cryo-EM) structures of sea urchin SLC9C1 in detergent and nanodiscs. We show that the voltage-sensing domains are positioned in an unusual configuration, sandwiching each side of the SLC9C1 homodimer. The S4 segment is very long, 90 Å in length, and connects the voltage-sensing domains to the cytoplasmic cyclic-nucleotide-binding domains. The S4 segment is in the up configuration-the inactive state of SLC9C1. Consistently, although a negatively charged cavity is accessible for Na⁺ to bind to the ion-transporting domains of SLC9C1, an intracellular helix connected to S4 restricts their movement. On the basis of the differences in the cryo-EM structure of SLC9C1 in the presence of cAMP, we propose that, upon hyperpolarization, the S4 segment moves down, removing this constriction and enabling Na⁺/H⁺ exchange.