Abstract
Vertebrate genomes encode 19 classical cadherins and about 100 nonclassical cadherins. Adhesion by classical cadherins depends on binding interactions in their N-terminal EC1 domains, which swap N-terminal β-strands between partner molecules from apposing cells. However, strand-swapping sequence signatures are absent from nonclassical cadherins, raising the question of how these proteins function in adhesion. Here, we show that T-cadherin, a glycosylphosphatidylinositol (GPI)-anchored cadherin, forms dimers through an alternative nonswapped interface near the EC1-EC2 calcium-binding sites. Mutations within this interface ablate the adhesive capacity of T-cadherin. These nonadhesive T-cadherin mutants also lose the ability to regulate neurite outgrowth from T-cadherin–expressing neurons. Our findings reveal the likely molecular architecture of the T-cadherin homophilic interface and its requirement for axon outgrowth regulation. The adhesive binding mode used by T-cadherin may also be used by other nonclassical cadherins.
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References
Gumbiner, B.M. Regulation of cadherin-mediated adhesion in morphogenesis. Nat. Rev. Mol. Cell Biol. 6, 622–634 (2005).
Takeichi, M. Morphogenetic roles of classic cadherins. Curr. Opin. Cell Biol. 7, 619–627 (1995).
Takeichi, M. The cadherin superfamily in neuronal connections and interactions. Nat. Rev. Neurosci. 8, 11–20 (2007).
Nollet, F., Kools, P. & van Roy, F. Phylogenetic analysis of the cadherin superfamily allows identification of six major subfamilies besides several solitary members. J. Mol. Biol. 299, 551–572 (2000).
Posy, S., Shapiro, L. & Honig, B. Sequence and structural determinants of strand swapping in cadherin domains: do all cadherins bind through the same adhesive interface? J. Mol. Biol. 378, 954–968 (2008).
Bekirov, I.H., Needleman, L.A., Zhang, W. & Benson, D.L. Identification and localization of multiple classic cadherins in developing rat limbic system. Neuroscience 115, 213–227 (2002).
Nishimura, E.K., Yoshida, H., Kunisada, T. & Nishikawa, S.I. Regulation of E- and P-cadherin expression correlated with melanocyte migration and diversification. Dev. Biol. 215, 155–166 (1999).
Price, S.R., De Marco Garcia, N.V., Ranscht, B. & Jessell, T.M. Regulation of motor neuron pool sorting by differential expression of type II cadherins. Cell 109, 205–216 (2002).
Wu, Q. & Maniatis, T. A striking organization of a large family of human neural cadherin-like cell adhesion genes. Cell 97, 779–790 (1999).
Usui, T. et al. Flamingo, a seven-pass transmembrane cadherin, regulates planar cell polarity under the control of Frizzled. Cell 98, 585–595 (1999).
Siemens, J. et al. Cadherin 23 is a component of the tip link in hair-cell stereocilia. Nature 428, 950–955 (2004).
Patel, S.D., Chen, C.P., Bahna, F., Honig, B. & Shapiro, L. Cadherin-mediated cell-cell adhesion: sticking together as a family. Curr. Opin. Struct. Biol. 13, 690–698 (2003).
Boggon, T.J. et al. C-cadherin ectodomain structure and implications for cell adhesion mechanisms. Science 296, 1308–1313 (2002).
Nagar, B., Overduin, M., Ikura, M. & Rini, J.M. Structural basis of calcium-induced E-cadherin rigidification and dimerization. Nature 380, 360–364 (1996).
Pokutta, S., Herrenknecht, K., Kemler, R. & Engel, J. Conformational changes of the recombinant extracellular domain of E-cadherin upon calcium binding. Eur. J. Biochem. 223, 1019–1026 (1994).
Goodwin, M. & Yap, A.S. Classical cadherin adhesion molecules: coordinating cell adhesion, signaling and the cytoskeleton. J. Mol. Histol. 35, 839–844 (2004).
Drees, F., Pokutta, S., Yamada, S., Nelson, W.J. & Weis, W.I. α-catenin is a molecular switch that binds E-cadherin-β-catenin and regulates actin-filament assembly. Cell 123, 903–915 (2005).
Haussinger, D. et al. Proteolytic E-cadherin activation followed by solution NMR and X-ray crystallography. EMBO J. 23, 1699–1708 (2004).
Pertz, O. et al. A new crystal structure, Ca2+ dependence and mutational analysis reveal molecular details of E-cadherin homoassociation. EMBO J. 18, 1738–1747 (1999).
Shapiro, L. et al. Structural basis of cell-cell adhesion by cadherins. Nature 374, 327–337 (1995).
Tamura, K., Shan, W.S., Hendrickson, W.A., Colman, D.R. & Shapiro, L. Structure-function analysis of cell adhesion by neural (N-) cadherin. Neuron 20, 1153–1163 (1998).
Patel, S.D. et al. Type II cadherin ectodomain structures: implications for classical cadherin specificity. Cell 124, 1255–1268 (2006).
Chen, C.P., Posy, S., Ben-Shaul, A., Shapiro, L. & Honig, B.H. Specificity of cell-cell adhesion by classical cadherins: critical role for low-affinity dimerization through β-strand swapping. Proc. Natl. Acad. Sci. USA 102, 8531–8536 (2005).
Ranscht, B. & Dours-Zimmermann, M.T. T-cadherin, a novel cadherin cell adhesion molecule in the nervous system lacks the conserved cytoplasmic region. Neuron 7, 391–402 (1991).
Vestal, D.J. & Ranscht, B. Glycosyl phosphatidylinositol–anchored T-cadherin mediates calcium-dependent, homophilic cell adhesion. J. Cell Biol. 119, 451–461 (1992).
Miskevich, F., Zhu, Y., Ranscht, B. & Sanes, J.R. Expression of multiple cadherins and catenins in the chick optic tectum. Mol. Cell. Neurosci. 12, 240–255 (1998).
Doyle, D.D. et al. T-cadherin is a major glycophosphoinositol-anchored protein associated with noncaveolar detergent-insoluble domains of the cardiac sarcolemma. J. Biol. Chem. 273, 6937–6943 (1998).
Koller, E. & Ranscht, B. Differential targeting of T- and N-cadherin in polarized epithelial cells. J. Biol. Chem. 271, 30061–30067 (1996).
Sacristan, M.P., Vestal, D.J., Dours-Zimmermann, M.T. & Ranscht, B. T-cadherin 2: molecular characterization, function in cell adhesion, and coexpression with T-cadherin and N-cadherin. J. Neurosci. Res. 34, 664–680 (1993).
Dames, S.A. et al. Insights into the low adhesive capacity of human T-cadherin from the NMR structure of its N-terminal extracellular domain. J. Biol. Chem. 283, 23485–23495 (2008).
Fredette, B.J. & Ranscht, B. T-cadherin expression delineates specific regions of the developing motor axon-hindlimb projection pathway. J. Neurosci. 14, 7331–7346 (1994).
Fredette, B.J., Miller, J. & Ranscht, B. Inhibition of motor axon growth by T-cadherin substrata. Development 122, 3163–3171 (1996).
Ivanov, D. et al. Expression of cell adhesion molecule T-cadherin in the human vasculature. Histochem. Cell Biol. 115, 231–242 (2001).
Hebbard, L.W. et al. T-cadherin supports angiogenesis and adiponectin association with the vasculature in a mouse mammary tumor model. Cancer Res. 68, 1407–1416 (2008).
Hug, C. et al. T-cadherin is a receptor for hexameric and high-molecular-weight forms of Acrp30/adiponectin. Proc. Natl. Acad. Sci. USA 101, 10308–10313 (2004).
Harrison, O. et al. Two-step adhesive binding by classical cadherins. Nat. Struct. Mol. Biol. advance online publication, doi:10:1038/nsmb.1784 (28 February 2010).
Harrison, O.J., Corps, E.M. & Kilshaw, P.J. Cadherin adhesion depends on a salt bridge at the N-terminus. J. Cell Sci. 118, 4123–4130 (2005).
Haussinger, D. et al. Calcium-dependent homoassociation of E-cadherin by NMR spectroscopy: changes in mobility, conformation and mapping of contact regions. J. Mol. Biol. 324, 823–839 (2002).
Chappuis-Flament, S., Wong, E., Hicks, L.D., Kay, C.M. & Gumbiner, B.M. Multiple cadherin extracellular repeats mediate homophilic binding and adhesion. J. Cell Biol. 154, 231–243 (2001).
Koch, A.W., Pokutta, S., Lustig, A. & Engel, J. Calcium binding and homoassociation of E-cadherin domains. Biochemistry 36, 7697–7705 (1997).
Bai, S., Datta, J., Jacob, S.T. & Ghoshal, K. Treatment of PC12 cells with nerve growth factor induces proteasomal degradation of T-cadherin that requires tyrosine phosphorylation of its cadherin domain. J. Biol. Chem. 282, 27171–27180 (2007).
Bai, S., Ghoshal, K. & Jacob, S.T. Identification of T-cadherin as a novel target of DNA methyltransferase 3B and its role in the suppression of nerve growth factor-mediated neurite outgrowth in PC12 cells. J. Biol. Chem. 281, 13604–13611 (2006).
Wichterle, H., Lieberam, I., Porter, J.A. & Jessell, T.M. Directed differentiation of embryonic stem cells into motor neurons. Cell 110, 385–397 (2002).
Parisini, E., Higgins, J.M., Liu, J.H., Brenner, M.B. & Wang, J.H. The crystal structure of human E-cadherin domains 1 and 2, and comparison with other cadherins in the context of adhesion mechanism. J. Mol. Biol. 373, 401–411 (2007).
Alattia, J.R. et al. Lateral self-assembly of E-cadherin directed by cooperative calcium binding. FEBS Lett. 417, 405–408 (1997).
Katsamba, P. et al. Linking molecular affinity and cellular specificity in cadherin-mediated adhesion. Proc. Natl. Acad. Sci. USA 106, 11594–11599 (2009).
Otwinowski, Z. & Minor, W. Processing of X-ray data collected in oscillation mode. Methods Enzymol. 276, 307–326 (1997).
Leslie, A.G.W. Recent changes to the MOSFLM package for processing film and image plate data. Joint CCP4 and ESF-EACBM Newsletters on Protein Crystallography 26 (1992).
Collaborative Computation Project, Number 4. The CCP4 suite: programs for protein crystallography. Acta Crystallogr. D Biol. Crystallogr. 50, 760–763 (1994).
Terwilliger, T.C. & Berendzen, J. Automated MAD and MIR structure solution. Acta Crystallogr. D Biol. Crystallogr. 55, 849–861 (1999).
Bricogne, G., Vonrhein, C., Flensburg, C., Schiltz, M. & Paciorek, W. Generation, representation and flow of phase information in structure determination: recent developments in and around SHARP 2.0. Acta Crystallogr. D Biol. Crystallogr. 59, 2023–2030 (2003).
Terwilliger, T. SOLVE and RESOLVE: automated structure solution, density modification and model building. J. Synchrotron Radiat. 11, 49–52 (2004).
Emsley, P. & Cowtan, K. Coot: model-building tools for molecular graphics. Acta Crystallogr. D Biol. Crystallogr. 60, 2126–2132 (2004).
Brunger, A.T. et al. Crystallography and NMR system (CNS): A new software system for macromolecular structure determination. Acta Crystallogr. D Biol. Crystallogr. 54, 905–921 (1998).
Murshudov, G.N., Vagin, A.A. & Dodson, E.J. Refinement of macromolecular structures by the maximum-likelihood method. Acta Crystallogr. D Biol. Crystallogr. 53, 240–255 (1997).
McCoy, A.J. et al. Phaser crystallographic software. J. Appl. Crystallogr. 40, 658–674 (2007).
Domeniconi, M. et al. MAG induces regulated intramembrane proteolysis of the p75 neurotrophin receptor to inhibit neurite outgrowth. Neuron 46, 849–855 (2005).
Acknowledgements
We thank P.D. Kwong for helpful suggestions on the manuscript. This work was supported in part by US National Institutes of Health grants R01 GM062270 (L.S.), U54 CA121852 (B.H. and L.S.) and R01 GM30518 (B.H.) and US National Science Foundation grants MCB-0416708 (B.H.), PO1 HD25938 (B.R.) and T32 GM08666 (H.C.V.). B.H. and T.M.J. are investigators of the Howard Hughes Medical Institute. X-ray data were acquired at the X4A and X4C beamlines of the National Synchrotron Light Source, Brookhaven National Laboratory; the X4 beamlines are operated by the New York Structural Biology Center. Use of the SGX Collaborative Access Team (SGX-CAT) beamline facilities at Sector 31 of the Advanced Photon Source was provided by SGX Pharmaceuticals, Inc., which constructed and operates the facility.
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C.C. determined and refined all the crystal structures; F.B. produced all the wild-type recombinant proteins; N.Z. performed the neurite outgrowth assays and surface biotinylation; H.C.V. performed the cell-aggregation studies; P.S.K. performed the SPR experiments; G.A. performed the AUC experiments; O.J.H. and J.B. prepared mutant proteins; X.J. helped in crystallographic data collection and refinement; S.P. and J.V. performed bioinformatic analysis; B.R. designed and analyzed cell-based experiments, T.M.J., B.H. and L.S. designed experiments, analyzed data, and wrote the manuscript.
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Ciatto, C., Bahna, F., Zampieri, N. et al. T-cadherin structures reveal a novel adhesive binding mechanism. Nat Struct Mol Biol 17, 339–347 (2010). https://doi.org/10.1038/nsmb.1781
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DOI: https://doi.org/10.1038/nsmb.1781
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