Peer-reviewed veterinary case report
Protocol for purification, single-particle cryo-EM structure determination, and functional assays of the fungal plasma membrane H<sup>+</sup>-ATPase.
- Year:
- 2025
- Authors:
- Ni YR et al.
- Affiliation:
- Department of Biophysics · China
Abstract
The fungal plasma membrane H<sup>+</sup>-ATPase (Pma1) maintains intracellular pH and membrane potential, but its study has been limited by challenges in sample preparation and functional assays. Here, we present a protocol for the endogenous purification of Pma1 in Saccharomyces cerevisiae, assessment of ATPase activity, cryo-electron microscopy (cryo-EM) structure determination, generation of a conditional pma1 knockout strain, and the performance of growth assays on mutants. For complete details on the use and execution of this protocol, please refer to You et al.<sup>1</sup>.
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Search related cases →Original publication: https://europepmc.org/article/MED/41343279