New Insights into the Structure-Function Relationship of the Endosomal-Type Na⁺, K⁺/H⁺ Antiporter NHX6 from Mulberry (<i>Morus notabilis</i>)

• Main Authors: Boning Cao, Zhongqiang Xia, Changying Liu, Wei Fan, Shuai Zhang, Qiao Liu, Zhonghuai Xiang, Aichun Zhao
• Format: Article
• Language: English
• Published: MDPI AG 2020-01-01
• Series: International Journal of Molecular Sciences
• Subjects: mulberry; na⁺, k⁺/h⁺ antiporters; salt-stress; homology modeling; ion transport
• Online Access: https://www.mdpi.com/1422-0067/21/2/428

The endosomal-type Na⁺, K⁺/H⁺ antiporters (NHXs) play important roles in K⁺, vesicle pH homeostasis, and protein trafficking in plant. However, the structure governing ion transport mechanism and the key residues related to the structure&#8722;function of the endosomal-type NHXs remain unclear. Here, the structure-function relationship of the only endosomal-type NHX from mulberry, MnNHX6, was investigated by homology modeling, mutagenesis, and localization analyses in yeast. The ectopic expression of MnNHX6 in arabidopsis and Nhx1 mutant yeast can enhance their salt tolerance. MnNHX6&#8217;s three-dimensional structure, established by homology modeling, was supported by empirical, phylogenetic, and experimental data. Structure analysis showed that MnNHX6 contains unusual 13 transmembrane helices, but the structural core formed by TM5-TM12 assembly is conserved. Localization analysis showed that MnNHX6 has the same endosomal localization as yeast Nhx1/VPS44, and Arg402 is important for protein stability of MnNHX6. Mutagenesis analysis demonstrated MnNHX6 contains a conserved cation binding mechanism and a similar charge-compensated pattern as NHE1, but shares a different role in ion selectivity than the vacuolar-type NHXs. These results improve our understanding of the role played by the structure&#8722;function related key residues of the plant endosomal-type NHXs, and provide a basis for the ion transport mechanism study of endosomal-type NHXs.