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Site–site communication in the EF-hand Ca2+-binding protein calbindin D9k

Nature Structural Biology volume 7, pages 245–250 (2000)Cite this article

Abstract

The cooperative binding of Ca2+ ions is an essential functional property of the EF-hand family of Ca2+-binding proteins. To understand how these proteins function, it is essential to characterize intermediate binding states in addition to the apo- and holo-proteins. The three-dimensional solution structure and fast time scale internal motional dynamics of the backbone have been determined for the half-saturated state of the N56A mutant of calbindin D9k with Ca2+ bound only in the N-terminal site. The extent of conformational reorganization and a loss of flexibility in the C-terminal EF-hand upon binding of an ion in the N-terminal EF-hand provide clear evidence of the importance of site–site interactions in this family of proteins, and demonstrates the strength of long-range effects in the cooperative EF-hand Ca2+-binding domain.

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Figure 1: Schematic diagram of the Ca2+-binding pathways in calbindin D9k.
Figure 2: Plots of a, per residue NOE-derived distance constraints and b, per residue r.m.s.d. from the mean structure for (Ca2+)1-N56A calbindin D9k.
Figure 3: Stereo view of the ensemble of 24 (Ca2+)1-N56A calbindin D9k structures.
Figure 4: Comparison of the packing of Phe 36 against side chains in helix IV in the apo state, (Ca2+)1-N56A, and (Ca2+)2 state of calbindin D9k.
Figure 5: Exchange terms for the two half-saturated states of calbindin D9k.
Figure 6: Comparison of backbone motional dynamics of different binding states of calbindin D9k.

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Acknowledgements

This research was supported by grants from the National Institutes of Health to W.J.C. and to M.R., and a postdoctoral fellowship (to L.M.) from the Swedish National Science Research Council. We thank J. Chung for assistance with NMR experiments and A.G. Palmer, III, for helpful discussions.

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Author notes

  1. Lena Mäler

    Present address: Department of Biophysics, Stockholm University, Stockholm, Sweden

Authors and Affiliations

  1. Department of Molecular Biology (MB9), The Scripps Research Institute, 10550 North Torrey Pines Road., La Jolla, 92037, California, USA

    Lena Mäler, John Blankenship & Walter J. Chazin

  2. Department of Molecular Genetics, Biochemistry, Microbiology, University of Cincinnati, Cincinnati, 45267, Ohio, USA

    Mark Rance

  3. Department of Biochemistry, Vanderbilt University, 606 Light Hall, Nashville, 37232-0146, Tennessee, USA

    Walter J. Chazin

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Correspondence to Walter J. Chazin.

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Mäler, L., Blankenship, J., Rance, M. et al. Site–site communication in the EF-hand Ca2+-binding protein calbindin D9k. Nat Struct Mol Biol 7, 245–250 (2000). https://doi.org/10.1038/73369

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