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Molecular mechanics of calcium–myristoyl switches

Nature volume 389, pages 198–202 (1997)Cite this article

Abstract

Many eukaryotic cellular and viral proteins have a covalently attached myristoyl group at the amino terminus. One such protein is recoverin, a calcium sensor in retinal rod cells, which controls the lifetime of photoexcited rhodopsin by inhibiting rhodopsin kinase1,2,3,4,5,6. Recoverin has a relative molecular mass of 23,000 (Mr 23K), and contains an amino-terminal myristoyl group (or related acyl group) and four EF hands7. The binding of two Ca2+ ions to recoverin leads to its translocation from the cytosol to the disc membrane8,9. In the Ca2+-free state, the myristoyl group is sequestered in a deep hydrophobic box, where it is clamped by multiple residues contributed by three of the EF hands10. We have used nuclear magnetic resonance to show that Ca2+ induces the unclamping and extrusion of the myristoyl group, enabling it to interact with a lipid bilayer membrane. The transition is also accompanied by a 45-degree rotation of the amino-terminal domain relative to the carboxy-terminal domain, and many hydrophobic residues are exposed. The conservation of the myristoyl binding site and two swivels in recoverin homologues from yeast to humans indicates that calcium–myristoyl switches are ancient devices for controlling calcium-sensitive processes.

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Figure 1: a, Superposition of the main-chain atoms of the 24 NMR-derived structures of myristoylated recoverin with two Ca2+ bound.
Figure 2: Space-filling model (a) and ribbon diagram (b) of Ca2+-free10 (left, liku.pdb) and Ca2+-bound (right) myristoylated recoverin.
Figure 3: Schematic ribbon representation of the structure of EF-2 and EF-3 of Ca2+-free (left) and Ca2+-bound (right) myristoylated recoverin.
Figure 4: Schematic ribbon representation of the structure of the N-terminal domain of Ca2+-free (top) and Ca2+-bound (bottom) recoverin.

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Acknowledgements

We thank G. Gokel for help with the synthesis of 13-oxatetradecanoic acid; L. Kay for help with NMR experiments; and F. Delaglio and D. Garrett for computer software for NMR data processing and analysis. This work was supported by grants to L.S. and J.G. from the NIH, and to M.I. from the Medical Research Council of Canada. J.B.A. was supported by an NIH post-doctoral fellowship. M.I. is a Howard Hughes Medical Institute international research scholar.

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Authors and Affiliations

  1. Department of Neurobiology, Stanford University School of Medicine, Stanford, 94305, California, USA

    James B. Ames & Lubert Stryer

  2. Division of Molecular and Structural Biology, Ontario Cancer Institute and Department of Medical Biophysics, University of Toronto, 610 University Avenue, Toronto, M5G 2M9, Ontario, Canada

    Rieko Ishima & Mitsuhiko Ikura

  3. Center for Tsukuba Advanced Research Alliance and Institute of Applied Biochemistry, University of Tsukuba, 305, Tsukuba, Japan

    Toshiyuki Tanaka & Mitsuhiko Ikura

  4. Department of Molecular Biology and Pharmacology, Washington University School of Medicine, St Louis, 63110, Missouri, USA

    Jeffrey I. Gordon

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Correspondence to Mitsuhiko Ikura.

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Ames, J., Ishima, R., Tanaka, T. et al. Molecular mechanics of calcium–myristoyl switches. Nature 389, 198–202 (1997). https://doi.org/10.1038/38310

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