Mechanosensitive （MS） channels are extensively stud- ied membrane protein for maintaining intracellular homeostasis through translocating solutes and ions across the membrane, but its mechanisms of channel gating an...Mechanosensitive （MS） channels are extensively stud- ied membrane protein for maintaining intracellular homeostasis through translocating solutes and ions across the membrane, but its mechanisms of channel gating and ion selectivity are largely unknown. Here, we identified the Ynal channel as the Na^＋/K^＋ cation-selec- tive MS channel and solved its structure at 3.8 A by cryo- EM single-particle method. Ynal exhibits low conduc- tance among the family of MS channels in E. coil, and shares a similar overall heptamer structure fold with previously studied MscS channels. By combining structural based mutagenesis, quantum mechanical and electrophysiological characterizations, we revealed that ion selective filter formed by seven hydrophobic methionine （Ynal^Met158） in the transmembrane pore determined ion selectivity, and both ion selectivity and gating of Ynal channel were affected by accompanying anions in solution. Further quantum simulation and functional validation support that the distinct binding energies with various anions to Ynal^Met158 facilitate Na^＋/K^＋ pass through, which was defined as binding-block mechanism. Our structural and functional studies provided a new perspective for understanding the mechanism of how MS channels select ions driven by mechanical force.展开更多
文摘Mechanosensitive （MS） channels are extensively stud- ied membrane protein for maintaining intracellular homeostasis through translocating solutes and ions across the membrane, but its mechanisms of channel gating and ion selectivity are largely unknown. Here, we identified the Ynal channel as the Na^＋/K^＋ cation-selec- tive MS channel and solved its structure at 3.8 A by cryo- EM single-particle method. Ynal exhibits low conduc- tance among the family of MS channels in E. coil, and shares a similar overall heptamer structure fold with previously studied MscS channels. By combining structural based mutagenesis, quantum mechanical and electrophysiological characterizations, we revealed that ion selective filter formed by seven hydrophobic methionine （Ynal^Met158） in the transmembrane pore determined ion selectivity, and both ion selectivity and gating of Ynal channel were affected by accompanying anions in solution. Further quantum simulation and functional validation support that the distinct binding energies with various anions to Ynal^Met158 facilitate Na^＋/K^＋ pass through, which was defined as binding-block mechanism. Our structural and functional studies provided a new perspective for understanding the mechanism of how MS channels select ions driven by mechanical force.
