6QK8

Crystal structure of yeast 14-3-3 protein (Bmh1) from Saccharomyces cerevisiae with the Nha1p (yeast Na+/H+ antiporter) 14-3-3 binding motif Ser481

6QK8 の概要

• エントリーDOI: 10.2210/pdb6qk8/pdb
• 分子名称: Protein BMH1, Na(+)/H(+) antiporter (2 entities in total)
• 機能のキーワード: 14-3-3 protein, bmh, na+/h+ antiporter, nha1p, signaling protein
• 由来する生物種: Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast); 詳細
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 111468.37

構造登録者

Smidova, A.,Obsil, T.,Obsilova, V. (登録日: 2019-01-28, 公開日: 2019-10-02, 最終更新日: 2024-11-20)

主引用文献

Smidova, A.,Stankova, K.,Petrvalska, O.,Lazar, J.,Sychrova, H.,Obsil, T.,Zimmermannova, O.,Obsilova, V.
The activity of Saccharomyces cerevisiae Na+, K+/H+antiporter Nha1 is negatively regulated by 14-3-3 protein binding at serine 481.
Biochim Biophys Acta Mol Cell Res, 1866:118534-118534, 2019

PubMed Abstract: Na/H antiporters are involved in ensuring optimal intracellular concentrations of alkali-metal cations and protons in most organisms. In Saccharomyces cerevisiae, the plasma-membrane Na, K/H antiporter Nha1 mediates Na and K efflux, which is important for cell growth in the presence of salts. Nha1 belongs among housekeeping proteins and, due to its ability to export K, it has many physiological functions. The Nha1 transport activity is regulated through its long, hydrophilic and unstructured C-terminus (554 of 985 aa). Although Nha1 has been previously shown to interact with the yeast 14-3-3 isoform (Bmh2), the binding site remains unknown. In this work, we identified the residues through which Nha1 interacts with the 14-3-3 protein. Biophysical characterization of the interaction between the C-terminal polypeptide of Nha1 and Bmh proteins in vitro revealed that the 14-3-3 protein binds to phosphorylated Ser481 of Nha1, and the crystal structure of the phosphopeptide containing Ser481 bound to Bmh1 provided the structural basis of this interaction. Our data indicate that 14-3-3 binding induces a disorder-to-order transition of the C-terminus of Nha1, and in vivo experiments showed that the mutation of Ser481 to Ala significantly increases cation efflux activity via Nha1, which renders cells sensitive to low K concentrations. Hence, 14-3-3 binding is apparently essential for the negative regulation of Nha1 activity, which should be low under standard growth conditions, when low amounts of toxic salts are present and yeast cells need to accumulate high amounts of K.

PubMed: 31446061
DOI: 10.1016/j.bbamcr.2019.118534

実験手法

X-RAY DIFFRACTION (2.917 Å)