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Nectin ectodomain structures reveal a canonical adhesive interface

Nature Structural & Molecular Biology volume 19, pages 906–915 (2012)Cite this article

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Abstract

Nectins are immunoglobulin superfamily glycoproteins that mediate intercellular adhesion in many vertebrate tissues. Homophilic and heterophilic interactions between nectin family members help mediate tissue patterning. We determined the homophilic binding affinities and heterophilic specificities of all four nectins and the related protein nectin-like 5 (Necl-5) from human and mouse, revealing a range of homophilic interaction strengths and a defined heterophilic specificity pattern. To understand the molecular basis of their adhesion and specificity, we determined the crystal structures of natively glycosylated full ectodomains or adhesive fragments of all four nectins and Necl-5. All of the crystal structures revealed dimeric nectins bound through a stereotyped interface that was previously proposed to represent a cis dimer. However, conservation of this interface and the results of targeted cross-linking experiments showed that this dimer probably represents the adhesive trans interaction. The structure of the dimer provides a simple molecular explanation for the adhesive binding specificity of nectins.

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Figure 1: SPR analysis of nectin binding interactions.
Figure 2: Overall structures of nectin homodimers.
Figure 3: Structural details of nectin homophilic dimers.
Figure 4: Targeted cross-linking of the nectin homodimer interface in transfected A431D cells.
Figure 5: Analysis of the molecular basis of nectin binding specificity.

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Acknowledgements

This work has been supported by grants from the US National Institutes of Health (AR44016 and AR057992 to S.M.T. and R01 GM062270 to L.S.) and from the National Science Foundation (MCB-0918535 to B.H.). Use of the Advanced Photon Source (APS) for data collection on human nectin-1 (D1–D3) and human nectin-4 (D1–D2) at beamline 24-ID-E was supported by the US Department of Energy, Office of Science, Office of Basic Energy Sciences under contract DE-AC02-06CH11357. X-ray data for all other nectins were acquired at the X4A and X4C beamlines of the National Synchrotron Light Source, Brookhaven National Laboratory (BNL); the beamlines are operated by the New York Structural Biology center. We thank J. Schwanof and R. Abramowitz at BNL and N. Sukumar at APS for support with synchrotron data collection.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biophysics, Columbia University, New York, New York, USA

    Oliver J Harrison, Jeremie Vendome, Julia Brasch, Xiangshu Jin, Phinikoula S Katsamba, Goran Ahlsen, Barry Honig & Lawrence Shapiro

  2. Howard Hughes Medical Institute, Columbia University, New York, New York, USA

    Oliver J Harrison, Jeremie Vendome, Xiangshu Jin, Phinikoula S Katsamba, Goran Ahlsen & Barry Honig

  3. Columbia Initiative in Systems Biology, Columbia University, New York, New York, USA

    Oliver J Harrison, Jeremie Vendome, Julia Brasch, Xiangshu Jin, Phinikoula S Katsamba, Goran Ahlsen, Barry Honig & Lawrence Shapiro

  4. Department of Dermatology, Northwestern University, The Feinberg School of Medicine, Chicago, Illinois, USA

    Soonjin Hong, Regina B Troyanovsky & Sergey M Troyanovsky

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Contributions

O.J.H., J.B. and X.J. determined and refined all crystal structures. O.J.H. produced all wild-type and mutant proteins. P.S.K. performed and analyzed the SPR experiments. G.A. performed and analyzed the AUC experiments. J.V. performed all bioinformatic analyses. S.H., R.B.T. and S.M.T. performed immunofluorescence and cross-linking studies. O.J.H., B.H. and L.S. designed experiments, analyzed data and wrote the manuscript.

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Correspondence to Barry Honig or Lawrence Shapiro.

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Harrison, O., Vendome, J., Brasch, J. et al. Nectin ectodomain structures reveal a canonical adhesive interface. Nat Struct Mol Biol 19, 906–915 (2012). https://doi.org/10.1038/nsmb.2366

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