Electrochemical coupling at the plasma membrane by mouse voltage-sensitive phosphatase requires association with basigin

Voltage-sensitive phosphatases (VSPs) are unique enzymes that mediate electrochemical coupling by converting phosphoinositides in response to membrane depolarization. In mammals, VSPs are involved in regulating sperm motility. The basic functionality of mammalian VSPs has remained enigmatic, as retention to intracellular compartments precluded functional analysis. Here, a membrane yeast two-hybrid assay identifies basigin as an accessory subunit of mouse VSP (mVSP). Co-expression with basigin or its homologs induced trafficking of mVSP from the endoplasmic reticulum to the plasma membrane. Mutational analysis and structural predictions by AlphaFold-Multimer showed that the functional effect on mVSP requires interaction of the transmembrane region of basigin with the voltage sensor domain of mVSP. Basigin-mediated surface localization allowed for functional analysis, revealing that mVSP acts as a voltage-activated 5-phosphatase against PI(4,5)P₂ and PI(3,4,5)P₃ with a more negative activation range compared to non-mammalian VSPs.