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Do's and Don'ts of Cryo-electron Microscopy: A Primer on Sample Preparation and High Quality Data Collection for Macromolecular 3D Reconstruction
Published on: January 9, 2015
Cryo-EM structure of human Na+/H+ exchanger NHA1 in an inward-open conformation
Bingjie Tang1, Qinling Qiu1, Shangyu Dang2
1Division of Life Science, The Hong Kong University of Science and Technology, Hong Kong, China.
Abstract:
The human Na+/H+ exchanger NHA1 (SLC9B1) plays a critical role in sperm motility and male fertility by regulating intracellular pH through its sodium-proton antiport activity. Despite its physiological importance, the structural and mechanistic basis of NHA1 function has remained elusive. Here, we present the cryo-EM structure of human NHA1 at 3.6 Å resolution, captured in an inward-open conformation. Structural comparisons with modeled outward-open states reveal substantial conformational changes in the core domain, supporting an elevator-like transport mechanism. Additionally, several key residues are identified that modify their interaction patterns, likely aiding in the conformational switches necessary for substrate transport. Their contribution to substrate transport is further supported by mutagenesis and sodium transport assays, providing insights into the molecular mechanism of NHA1.
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