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Solution structures of the adhesion molecule DdCAD-1 reveal new insights into Ca2+-dependent cell-cell adhesion

Nature Structural & Molecular Biology volume 13, pages 10161022 (2006) | Download Citation

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Abstract

DdCAD-1 is a novel Ca2+-dependent cell adhesion molecule that lacks a hydrophobic signal peptide and a transmembrane domain. DdCAD-1 is expressed by the social amoeba Dictyostelium discoideum at the onset of development. It is synthesized as a soluble protein and then transported to the plasma membrane by contractile vacuoles. Here we describe the novel features of the solution structures of Ca2+-free and Ca2+-bound monomeric DdCAD-1. DdCAD-1 contains two β-sandwich domains, belonging to the βγ-crystallin and immunoglobulin fold classes, respectively. Whereas the N-terminal domain has a major role in homophilic binding, the C-terminal domain tethers the protein to the cell membrane. From structural and mutational analyses, we propose a model for the Ca2+-bound DdCAD-1 dimer as a basis for understanding DdCAD-1–mediated cell-cell adhesion at the molecular level. Our results provide new insights into Ca2+-dependent mechanisms for cell-cell adhesion.

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Acknowledgements

We thank J.S. Fan for assistance in NMR experiments and Y.K. Mok (National University of Singapore) for providing a pET32a-derived expression vector used in this work. This research was supported by a research grant from the National University of Singapore to D.W.Y. and by Operating Grant MT-6140 from the Canadian Institutes of Health Research to C.-H.S.

Author information

Affiliations

  1. Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543.

    • Zhi Lin
    •  & Daiwen Yang
  2. Banting and Best Department of Medical Research and Department of Biochemistry, University of Toronto, Toronto, Ontario M5G 1L6, Canada.

    • Shrivani Sriskanthadevan
    • , Haibo Huang
    •  & Chi-Hung Siu

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Contributions

Z.L. contributed to NMR sample preparations, assignments, structure calculations, mutagenesis studies, model construction, in vitro homoassociation studies and manuscript preparation; S.S. and H.H. contributed to molecular cloning, mutagenesis, protein expression and binding assays in subdomain studies; C.-H.S contributed to project guidance and manuscript preparation; and D.W.Y. contributed to project guidance, NMR data collection and manuscript preparation.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Chi-Hung Siu or Daiwen Yang.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    Effects of Ca2+ and Mn2+ on 1H-15N HSQC spectra of DdCAD-1.

  2. 2.

    Supplementary Fig. 2

    Chemical shift changes induced by Ca2+ binding to DdCAD-1.

  3. 3.

    Supplementary Fig. 3

    1H or 15N chemical shift titration curves for the binding of Ca2+ to DdCAD-1.

  4. 4.

    Supplementary Fig. 4

    Structural comparisons of DdCAD-1 with protein S and cadherin.

  5. 5.

    Supplementary Fig. 5

    Structure-based sequence alignments.

  6. 6.

    Supplementary Fig. 6

    Homophilic interaction of DdCAD-1 in vitro.

  7. 7.

    Supplementary Fig. 7

    CD spectra of DdCAD-1 mutants.

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    Supplementary Fig. 8

    Binding of 45Ca2+ to wild-type and mutant His6–DdCAD-1 proteins.

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    Supplementary Table 1

    Statistics of alignments among DdCAD-1, protein S and cadherins.

  10. 10.

    Supplementary Table 2

    Structural statistics for the 10 lowest-energy Ca2+-bound DdCAD-1 dimers.

  11. 11.

    Supplementary Methods

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DOI

https://doi.org/10.1038/nsmb1162

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