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Journal of Biomolecular NMR

, Volume 71, Issue 2, pp 91–100 | Cite as

Binding of a small molecule water channel inhibitor to aquaporin Z examined by solid-state MAS NMR

Article

Abstract

Aquaporins are integral membrane proteins that facilitate water flow across biological membranes. Their involvement in multiple physiological functions and disease states has prompted intense research to discover water channel activity modulators. However, inhibitors found so far are weak and/or lack specificity. For organic compounds, which lack of high electron-dense atoms, the identification of binding sites is even more difficult. Nuclear magnetic resonance spectroscopy (NMR) requires large amounts of the protein, and expression and purification of mammalian aquaporins in large quantities is a difficult task. However, since aquaporin Z (AqpZ) can be purified and expressed in good quantities and has a high similarity to human AQP1 (~ 40% identity), it can be used as a model for studying the structure and function of human aquaporins. In the present study, we have used solid-state MAS NMR to investigate the binding of a lead compound [1-(4-methylphenyl)1H-pyrrole-2,5-dione] to AqpZ, through mapping of chemical shift perturbations in the presence of the compound.

Keywords

Solid-state nuclear magnetic resonance Membrane protein Aquaporin Z Liposomes Binding site Water channel inhibition 

Notes

Acknowledgements

This work was supported by Singapore National Research Foundation (NRF) Competitive Research Programme (CRP) NRF-CRP4-2008-02, and MOE Singapore Tier 1 Grant RG150/14. The authors acknowledge the graduate scholarship provided by the Singapore MOE. All solid-state MAS NMR experiments were performed by Dr. Nagashima and Toshio Yamazaki.

Supplementary material

10858_2018_195_MOESM1_ESM.docx (656 kb)
Supplementary material 1 (DOCX 656 KB)

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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.School of Biological SciencesNanyang Technological UniversitySingaporeSingapore
  2. 2.RIKEN Centre for Life Science TechnologiesKanagawaJapan

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