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A novel target recognition revealed by calmodulin in complex with Ca2+-calmodulin-dependent kinase kinase

Nature Structural Biology volume 6, pages 819–824 (1999)Cite this article

Abstract

The structure of calcium-bound calmodulin (Ca2+/CaM) complexed with a 26-residue peptide, corresponding to the CaM-binding domain of rat Ca2+/CaM-dependent protein kinase kinase (CaMKK), has been determined by NMR spectroscopy. In this complex, the CaMKK peptide forms a fold comprising an α-helix and a hairpin-like loop whose C-terminus folds back on itself. The binding orientation of this CaMKK peptide by the two CaM domains is opposite to that observed in all other CaM–target complexes determined so far. The N- and C-terminal hydrophobic pockets of Ca2+/CaM anchor Trp 444 and Phe 459 of the CaMKK peptide, respectively. This 14-residue separation between two key hydrophobic groups is also unique among previously determined CaM complexes. The present structure represents a new and distinct class of Ca2+/CaM target recognition that may be shared by other Ca2+/CaM-stimulated proteins.

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Figure 1: a, Alignment of Ca 2+ /CaM binding sequences of CaMKK, MLCK and CaMKII.
Figure 2: a, Schematic drawing of interacting residues between Ca 2+ /CaM and CaMKK peptide.
Figure 3: Electrostatic potential surfaces of the CaM–target peptide complexes.
Figure 4: Alignment of the sequences of CaM-binding region based on the position of the N-terminal key residue.

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Acknowledgements

We are grateful to C. Klee for kindly providing the Xenopus calmodulin expression system, K.I. Tong for calmodulin purification, N. Takahashi for the mutagenesis experiments, R. Ishima and T. Tanaka for NMR experiments and discussions. This work was in part supported by a grant (to M.I.) from Medical Research Council of Canada (MRCC). M.I. is a Howard Hughes Medical Institute International Research Scholar and MRCC Scientist.

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Author notes

  1. Mark B. Swindells

    Present address: Inpharmatica, London, W1P 2AX, UK

Authors and Affiliations

  1. Molecular Chemistry Research, Chemistry Laboratories, Institute for Drug Discovery Research, Yamanouchi Pharmaceutical Co., Ltd., Tsukuba, 305-8585, Japan

    Masanori Osawa, Hiroyuki Kurihara, Masaya Orita & Tadao Shibanuma

  2. Center for Tsukuba Advanced Research Alliance and Institute of Applied Biochemistry, University of Tsukuba, Tsukuba, 305-0006, Japan

    Masanori Osawa & Mark B. Swindells

  3. Helix Research Institute, Kisarazu, 292-0812, Japan

    Hiroshi Tokumitsu, Mark B. Swindells & Toshio Furuya

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

    Mitsuhiko Ikura

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Osawa, M., Tokumitsu, H., Swindells, M. et al. A novel target recognition revealed by calmodulin in complex with Ca2+-calmodulin-dependent kinase kinase. Nat Struct Mol Biol 6, 819–824 (1999). https://doi.org/10.1038/12271

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