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Synthetic partial agonists reveal key steps in IP3 receptor activation

Nature Chemical Biology volume 5, pages 631–639 (2009)Cite this article

Abstract

Inositol 1,4,5-trisphosphate receptors (IP3Rs) are ubiquitous intracellular Ca2+ channels. IP3 binding to the IP3-binding core (IBC) near the N terminus initiates conformational changes that lead to opening of a pore. The mechanisms underlying this process are unresolved. We synthesized 2-O–modified IP3 analogs that are partial agonists of IP3R. These are similar to IP3 in their interactions with the IBC, but they are less effective than IP3 in rearranging the relationship between the IBC and the N-terminal suppressor domain (SD), and they open the channel at slower rates. IP3R with a mutation in the SD occupying a position similar to the 2-O substituent of the partial agonists has a reduced open probability that is similar for full and partial agonists. Bulky or charged substituents from either the ligand or the SD therefore block obligatory coupling of the IBC and the SD. Analysis of ΔG for ligand binding shows that IP3 is recognized by the IBC and conformational changes then propagate entirely via the SD to the pore.

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Figure 1: Structure of the IP3R and its ligands.
Figure 2: 2-O–modified IP3 analogs are partial agonists of IP3R.
Figure 3: Partial agonists are similar to IP3 in their interactions with the IBC.
Figure 4: Point mutations within the SD mimic partial agonists.
Figure 5: IP3 binding to the IBC activates IP3R entirely via the SD.

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Acknowledgements

We thank S. Dedos, P. da Fonseca, A. Burgen, S. Otto and M. Garcia Alai for helpful comments, and T. Woodman for advice on NMR spectroscopy. This work was supported by grants from the Wellcome Trust (to C.W.T., A.M. Riley and B.V.L.P.) and the Biotechnology and Biological Sciences Research Council (to C.W.T.). A.M. Rossi holds a Junior Research Fellowship at Queens' College, Cambridge, UK.

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Author notes

  1. Ana M Rossi and Andrew M Riley: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pharmacology, University of Cambridge, Cambridge, UK

    Ana M Rossi, Stephen C Tovey, Taufiq Rahman, Olivier Dellis, Emily J A Taylor & Colin W Taylor

  2. Department of Pharmacy and Pharmacology, Wolfson Laboratory of Medicinal Chemistry, University of Bath, Claverton Down, Bath, UK

    Andrew M Riley & Barry V L Potter

  3. Department of Cell Biophysics, Institute of Biophysics and Cell Engineering, National Academy of Sciences of Belarus, Minsk, Belarus

    Valery G Veresov

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Contributions

A.M. Rossi, S.C.T., T.R., O.D. and E.J.A.T. completed the biology experiments. V.G.V. performed molecular modeling. A.M. Riley designed and synthesized the ligands and contributed to molecular modeling. B.V.L.P. (chemistry) and C.W.T. (biology) designed and coordinated the project. C.W.T. and A.M. Rossi wrote the manuscript with input from the other authors. All authors discussed the results and commented on the manuscript.

Corresponding authors

Correspondence to Barry V L Potter or Colin W Taylor.

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Rossi, A., Riley, A., Tovey, S. et al. Synthetic partial agonists reveal key steps in IP3 receptor activation. Nat Chem Biol 5, 631–639 (2009). https://doi.org/10.1038/nchembio.195

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