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Proton pump inhibitors have pH-dependent effects on the thermostability of the carboxyl-terminal domain of voltage-gated proton channel Hv1

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Abstract

The voltage-gated proton channel Hv1 is highly selective for H+ and is activated by membrane depolarization and pH gradient. An increased external and decreased internal pH opens the Hv1 channel. The intracellular C-terminal domain of Hv1 is responsible for channel dimerization, cooperative, and thermosensitive gating. Here, we found that proton pump inhibitors (PPIs) interact with the C-terminal domain of human Hv1. The interaction between PPIs and the C-terminal domain, which is pH-dependent, lowered the thermal and structural stability of the protein at pH 4, but enhanced the thermal and structural stability at pH 8. Furthermore, we investigated in vitro the interaction of PPIs with the C-terminal domain of Hv1 by fluorescence and micro-Raman spectra. Fluorescence quenching measurements revealed that the interaction between the C-terminal domain and PPIs is a mainly hydrophobic interaction. The micro-Raman spectra showed that PPIs did not form stable disulfide bonds with the unique thiol group within this domain (Cys249 residue). The preferential interaction of PPIs with the inactive form of Hv1 stabilizes the high pH inactive state of the C-terminal domain, indicating a mechanism by which PPIs might act explicitly on the stabilization of a closed state of the proton channel.

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Acknowledgements

This work was supported by National Natural Science Foundation of China (Nos. 30970579 and 31271464), the Ph.D. Programs Foundation of Ministry of Education of China (Nos. 20110031110004 and 20120031110028), the Basic Science and Advance Technology Research Program of Tianjin (No. 14JCYBJC23400), and the China Postdoctoral Science Foundation Funded Project (No. 2015M581289). This research is based on the Cooperative Research Project of Research Institute of Electronics, Shizuoka University.

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  1. Department of Biophysics, The Key Laboratory of Bioactive Materials, Ministry of Education, School of Physics Science, Nankai University, 94 Weijin Road, Nankai District, Tianjin, 300071, People’s Republic of China

    Qing Zhao, Weiyan Zuo, Shangrong Zhang, Yongqiang Zhang, Chuanyong Li & Shu Jie Li

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  1. Qing Zhao
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  2. Weiyan Zuo
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Correspondence to Shu Jie Li.

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Zhao, Q., Zuo, W., Zhang, S. et al. Proton pump inhibitors have pH-dependent effects on the thermostability of the carboxyl-terminal domain of voltage-gated proton channel Hv1. Eur Biophys J 47, 237–247 (2018). https://doi.org/10.1007/s00249-017-1253-3

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