Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Cryo-EM structure of the exocyst complex

An Author Correction to this article was published on 05 November 2018

This article has been updated


The exocyst is an evolutionarily conserved octameric protein complex that mediates the tethering of post-Golgi secretory vesicles to the plasma membrane during exocytosis and is implicated in many cellular processes such as cell polarization, cytokinesis, ciliogenesis and tumor invasion. Using cryo-EM and chemical cross-linking MS (CXMS), we solved the structure of the Saccharomyces cerevisiae exocyst complex at an average resolution of 4.4 Å. Our model revealed the architecture of the exocyst and led to the identification of the helical bundles that mediate the assembly of the complex at its core. Sequence analysis suggests that these regions are evolutionarily conserved across eukaryotic systems. Additional cell biological data suggest a mechanism for exocyst assembly that leads to vesicle tethering at the plasma membrane.

This is a preview of subscription content

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Fig. 1: Cryo-EM structure of the exocyst complex.
Fig. 2: The structure of the holo-exocyst complex and individual subunits.
Fig. 3: Pairwise interactions of the exocyst subunits through the CorEx motifs.
Fig. 4: Assembly of the core subcomplexes and the holocomplex.
Fig. 5: The CorEx motif of Sec3 is crucial for exocyst complex assembly and vesicle tethering.
Fig. 6: The SEC3 CorEx deletion mutant is defective in exocytosis.

Change history

  • 05 November 2018

    In the version of this article originally published, the value given for electron dose in Table 1 was incorrect. This value was originally stated as 4.8 but should have been 50. The error has been corrected in the HTML and PDF versions of the article.


  1. 1.

    Pfeffer, S. R. Transport-vesicle targeting: tethers before SNAREs. Nat. Cell. Biol. 1, E17–E22 (1999).

    CAS  Article  Google Scholar 

  2. 2.

    Guo, W., Sacher, M., Barrowman, J., Ferro-Novick, S. & Novick, P. Protein complexes in transport vesicle targeting. Trends Cell. Biol. 10, 251–255 (2000).

    CAS  Article  Google Scholar 

  3. 3.

    Whyte, J. R. & Munro, S. Vesicle tethering complexes in membrane traffic. J. Cell. Sci. 115, 2627–2637 (2002).

    CAS  PubMed  Google Scholar 

  4. 4.

    Sztul, E. & Lupashin, V. Role of tethering factors in secretory membrane traffic. Am. J. Physiol. Cell. Physiol. 290, C11–C26 (2006).

    CAS  Article  Google Scholar 

  5. 5.

    Yu, I. M. & Hughson, F. M. Tethering factors as organizers of intracellular vesicular traffic. Annu. Rev. Cell. Dev. Biol. 26, 137–156 (2010).

    CAS  Article  Google Scholar 

  6. 6.

    Bröcker, C., Engelbrecht-Vandré, S. & Ungermann, C. Multisubunit tethering complexes and their role in membrane fusion. Curr. Biol. 20, R943–R952 (2010).

    Article  Google Scholar 

  7. 7.

    TerBush, D. R. & Novick, P. Sec6, Sec8, and Sec15 are components of a multisubunit complex which localizes to small bud tips in Saccharomyces cerevisiae. J. Cell. Biol. 130, 299–312 (1995).

    CAS  Article  Google Scholar 

  8. 8.

    TerBush, D. R., Maurice, T., Roth, D. & Novick, P. The Exocyst is a multiprotein complex required for exocytosis in Saccharomyces cerevisiae. EMBO J. 15, 6483–6494 (1996).

    CAS  Article  Google Scholar 

  9. 9.

    He, B. & Guo, W. The exocyst complex in polarized exocytosis. Curr. Opin. Cell. Biol. 21, 537–542 (2009).

    CAS  Article  Google Scholar 

  10. 10.

    Wu, B. & Guo, W. The exocyst at a glance. J. Cell. Sci. 128, 2957–2964 (2015).

    CAS  Article  Google Scholar 

  11. 11.

    Yue, P. et al. Sec3 promotes the initial binary t-SNARE complex assembly and membrane fusion. Nat. Commun. 8, 14236 (2017).

    CAS  Article  Google Scholar 

  12. 12.

    Heider, M. R. & Munson, M. Exorcising the exocyst complex. Traffic 13, 898–907 (2012).

    CAS  Article  Google Scholar 

  13. 13.

    Munson, M. & Novick, P. The exocyst defrocked, a framework of rods revealed. Nat. Struct. Mol. Biol. 13, 577–581 (2006).

    CAS  Article  Google Scholar 

  14. 14.

    Hsu, S. C., Hazuka, C. D., Foletti, D. L. & Scheller, R. H. Targeting vesicles to specific sites on the plasma membrane: the role of the sec6/8 complex. Trends Cell. Biol. 9, 150–153 (1999).

    CAS  Article  Google Scholar 

  15. 15.

    Heider, M. R. et al. Subunit connectivity, assembly determinants and architecture of the yeast exocyst complex. Nat. Struct. Mol. Biol. 23, 59–66 (2016).

    CAS  Article  Google Scholar 

  16. 16.

    Lees, J. A., Yip, C. K., Walz, T. & Hughson, F. M. Molecular organization of the COG vesicle tethering complex. Nat. Struct. Mol. Biol. 17, 1292–1297 (2010).

    CAS  Article  Google Scholar 

  17. 17.

    Chou, H. T., Dukovski, D., Chambers, M. G., Reinisch, K. M. & Walz, T. CATCHR, HOPS and CORVET tethering complexes share a similar architecture. Nat. Struct. Mol. Biol. 23, 761–763 (2016).

    CAS  Article  Google Scholar 

  18. 18.

    Picco, A. et al. The in vivo architecture of the exocyst provides structural basis for exocytosis. Cell 168, 400–412.e18 (2017).

    CAS  Article  Google Scholar 

  19. 19.

    Ding, Y. H. et al. Increasing the depth of mass-spectrometry-based structural analysis of protein complexes through the use of multiple cross-linkers. Anal. Chem. 88, 4461–4469 (2016).

    CAS  Article  Google Scholar 

  20. 20.

    Croteau, N. J., Furgason, M. L., Devos, D. & Munson, M. Conservation of helical bundle structure between the exocyst subunits. PLoS. One 4, e4443 (2009).

    Article  Google Scholar 

  21. 21.

    Jackson, L. P., Kümmel, D., Reinisch, K. M. & Owen, D. J. Structures and mechanisms of vesicle coat components and multisubunit tethering complexes. Curr. Opin. Cell. Biol. 24, 475–483 (2012).

    CAS  Article  Google Scholar 

  22. 22.

    Sutton, R. B., Fasshauer, D., Jahn, R. & Brunger, A. T. Crystal structure of a SNARE complex involved in synaptic exocytosis at 2.4 A resolution. Nature 395, 347–353 (1998).

    CAS  Article  Google Scholar 

  23. 23.

    Finger, F. P., Hughes, T. E. & Novick, P. Sec3p is a spatial landmark for polarized secretion in budding yeast. Cell 92, 559–571 (1998).

    CAS  Article  Google Scholar 

  24. 24.

    Guo, W., Tamanoi, F. & Novick, P. Spatial regulation of the exocyst complex by Rho1 GTPase. Nat. Cell. Biol. 3, 353–360 (2001).

    CAS  Article  Google Scholar 

  25. 25.

    Zhang, X. et al. Membrane association and functional regulation of Sec3 by phospholipids and Cdc42. J. Cell. Biol. 180, 145–158 (2008).

    CAS  Article  Google Scholar 

  26. 26.

    Yamashita, M. et al. Structural basis for the Rho- and phosphoinositide-dependent localization of the exocyst subunit Sec3. Nat. Struct. Mol. Biol. 17, 180–186 (2010).

    CAS  Article  Google Scholar 

  27. 27.

    Baek, K. et al. Structure-function study of the N-terminal domain of exocyst subunit Sec3. J. Biol. Chem. 285, 10424–10433 (2010).

    CAS  Article  Google Scholar 

  28. 28.

    Boyd, C., Hughes, T., Pypaert, M. & Novick, P. Vesicles carry most exocyst subunits to exocytic sites marked by the remaining two subunits, Sec3p and Exo70p. J. Cell. Biol. 167, 889–901 (2004).

    CAS  Article  Google Scholar 

  29. 29.

    Luo, G., Zhang, J. & Guo, W. The role of Sec3p in secretory vesicle targeting and exocyst complex assembly. Mol. Biol. Cell. 25, 3813–3822 (2014).

    Article  Google Scholar 

  30. 30.

    He, B., Xi, F., Zhang, X., Zhang, J. & Guo, W. Exo70 interacts with phospholipids and mediates the targeting of the exocyst to the plasma membrane. EMBO J. 26, 4053–4065 (2007).

    CAS  Article  Google Scholar 

  31. 31.

    Liu, J., Zuo, X., Yue, P. & Guo, W. Phosphatidylinositol 4,5-bisphosphate mediates the targeting of the exocyst to the plasma membrane for exocytosis in mammalian cells. Mol. Biol. Cell. 18, 4483–4492 (2007).

    CAS  Article  Google Scholar 

  32. 32.

    Sivaram, M. V., Furgason, M. L., Brewer, D. N. & Munson, M. The structure of the exocyst subunit Sec6p defines a conserved architecture with diverse roles. Nat. Struct. Mol. Biol. 13, 555–556 (2006).

    CAS  Article  Google Scholar 

  33. 33.

    Jin, R. et al. Exo84 and Sec5 are competitive regulatory Sec6/8 effectors to the RalA GTPase. EMBO J. 24, 2064–2074 (2005).

    CAS  Article  Google Scholar 

  34. 34.

    Fukai, S., Matern, H. T., Jagath, J. R., Scheller, R. H. & Brunger, A. T. Structural basis of the interaction between RalA and Sec5, a subunit of the sec6/8 complex. EMBO J. 22, 3267–3278 (2003).

    CAS  Article  Google Scholar 

  35. 35.

    Wu, S., Mehta, S. Q., Pichaud, F., Bellen, H. J. & Quiocho, F. A. Sec15 interacts with Rab11 via a novel domain and affects Rab11 localization in vivo. Nat. Struct. Mol. Biol. 12, 879–885 (2005).

    CAS  Article  Google Scholar 

  36. 36.

    Dong, G., Hutagalung, A. H., Fu, C., Novick, P. & Reinisch, K. M. The structures of exocyst subunit Exo70p and the Exo84p C-terminal domains reveal a common motif. Nat. Struct. Mol. Biol. 12, 1094–1100 (2005).

    CAS  Article  Google Scholar 

  37. 37.

    Hamburger, Z. A., Hamburger, A. E., West, A. P. Jr. & Weis, W. I. Crystal structure of the S. cerevisiae exocyst component Exo70p. J. Mol. Biol. 356, 9–21 (2006).

    CAS  Article  Google Scholar 

  38. 38.

    Moore, B. A., Robinson, H. H. & Xu, Z. The crystal structure of mouse Exo70 reveals unique features of the mammalian exocyst. J. Mol. Biol. 371, 410–421 (2007).

    CAS  Article  Google Scholar 

  39. 39.

    Zhang, C. et al. Endosidin2 targets conserved exocyst complex subunit EXO70 to inhibit exocytosis. Proc. Natl Acad. Sci. USA 113, E41–E50 (2016).

    CAS  Article  Google Scholar 

  40. 40.

    Chen, J. et al. Crystal structure of Sec10, a subunit of the exocyst complex. Sci. Rep. 7, 40909 (2017).

    CAS  Article  Google Scholar 

  41. 41.

    Whyte, J. R. & Munro, S. The Sec34/35 Golgi transport complex is related to the exocyst, defining a family of complexes involved in multiple steps of membrane traffic. Dev. Cell. 1, 527–537 (2001).

    CAS  Article  Google Scholar 

  42. 42.

    Murray, D. H. et al. An endosomal tether undergoes an entropic collapse to bring vesicles together. Nature 537, 107–111 (2016).

    CAS  Article  Google Scholar 

  43. 43.

    Novick, P. & Guo, W. Ras family therapy: Rab, Rho and Ral talk to the exocyst. Trends Cell. Biol. 12, 247–249 (2002).

    CAS  Article  Google Scholar 

  44. 44.

    Lipschutz, J. H. & Mostov, K. E. Exocytosis: the many masters of the exocyst. Curr. Biol. 12, R212–R214 (2002).

    CAS  Article  Google Scholar 

  45. 45.

    Moskalenko, S. et al. The exocyst is a Ral effector complex. Nat. Cell. Biol. 4, 66–72 (2002).

    CAS  Article  Google Scholar 

  46. 46.

    Moskalenko, S. et al. Ral GTPases regulate exocyst assembly through dual subunit interactions. J. Biol. Chem. 278, 51743–51748 (2003).

    CAS  Article  Google Scholar 

  47. 47.

    Sugihara, K. et al. The exocyst complex binds the small GTPase RalA to mediate filopodia formation. Nat. Cell. Biol. 4, 73–78 (2002).

    CAS  Article  Google Scholar 

  48. 48.

    Luo, G., Zhang, J., Luca, F. C. & Guo, W. Mitotic phosphorylation of Exo84 disrupts exocyst assembly and arrests cell growth. J. Cell. Biol. 202, 97–111 (2013).

    CAS  Article  Google Scholar 

  49. 49.

    Lu, H. et al. Oncogenic BRAF-mediated melanoma cell invasion. Cell. Rep. 15, 2012–2024 (2016).

    CAS  Article  Google Scholar 

  50. 50.

    Ren, J. & Guo, W. ERK1/2 regulate exocytosis through direct phosphorylation of the exocyst component Exo70. Dev. Cell. 22, 967–978 (2012).

    CAS  Article  Google Scholar 

  51. 51.

    Yang, B. et al. Identification of cross-linked peptides from complex samples. Nat. Methods 9, 904–906 (2012).

    CAS  Article  Google Scholar 

  52. 52.

    Liu, X. & Wang, H. W. Single particle electron microscopy reconstruction of the exosome complex using the random conical tilt method. J. Vis. Exp. 49, 2574 (2011).

    Google Scholar 

  53. 53.

    Li, X., Zheng, S., Agard, D. A. & Cheng, Y. Aysnchronous data acquisition and on-the-fly analysis of dose fractionated cryo-EM images by UCSFImage. J. Struct. Biol. 192, 74–78 (2015).

    Article  Google Scholar 

  54. 54.

    Li, X. et al. Electron counting and beam-induced motion correction enable near-atomic-resolution single-particle cryo-EM. Nat. Methods 10, 584–590 (2013).

    CAS  Article  Google Scholar 

  55. 55.

    Scheres, S. H. Beam-induced motion correction for sub-megadalton cryo-EM particles. eLife 3, e03665 (2014).

    Article  Google Scholar 

  56. 56.

    Tang, G. et al. EMAN2: an extensible image processing suite for electron microscopy. J. Struct. Biol. 157, 38–46 (2007).

    CAS  Article  Google Scholar 

  57. 57.

    Shaikh, T. R. et al. SPIDER image processing for single-particle reconstruction of biological macromolecules from electron micrographs. Nat. Protoc. 3, 1941–1974 (2008).

    CAS  Article  Google Scholar 

  58. 58.

    Mindell, J. A. & Grigorieff, N. Accurate determination of local defocus and specimen tilt in electron microscopy. J. Struct. Biol. 142, 334–347 (2003).

    Article  Google Scholar 

  59. 59.

    Scheres, S. H. RELION: implementation of a Bayesian approach to cryo-EM structure determination. J. Struct. Biol. 180, 519–530 (2012).

    CAS  Article  Google Scholar 

  60. 60.

    Zheng, S. Q. et al. MotionCor2: anisotropic correction of beam-induced motion for improved cryo-electron microscopy. Nat. Methods 14, 331–332 (2017).

    CAS  Article  Google Scholar 

  61. 61.

    Grant, T. & Grigorieff, N. Automatic estimation and correction of anisotropic magnification distortion in electron microscopes. J. Struct. Biol. 192, 204–208 (2015).

    Article  Google Scholar 

  62. 62.

    Yu, G. et al. An algorithm for estimation and correction of anisotropic magnification distortion of cryo-EM images without need of pre-calibration. J. Struct. Biol. 195, 207–215 (2016).

    Article  Google Scholar 

  63. 63.

    Pettersen, E. F. et al. UCSF Chimera—a visualization system for exploratory research and analysis. J. Comput. Chem. 25, 1605–1612 (2004).

    CAS  Article  Google Scholar 

  64. 64.

    Kucukelbir, A., Sigworth, F. J. & Tagare, H. D. Quantifying the local resolution of cryo-EM density maps. Nat. Methods 11, 63–65 (2014).

    CAS  Article  Google Scholar 

  65. 65.

    Scheres, S. H. & Chen, S. Prevention of overfitting in cryo-EM structure determination. Nat. Methods 9, 853–854 (2012).

    CAS  Article  Google Scholar 

  66. 66.

    Wriggers, W., Milligan, R. A. & McCammon, J. A. Situs: a package for docking crystal structures into low-resolution maps from electron microscopy. J. Struct. Biol. 125, 185–195 (1999).

    CAS  Article  Google Scholar 

  67. 67.

    Biasini, M. et al. SWISS-MODEL: modelling protein tertiary and quaternary structure using evolutionary information. Nucleic Acids Res. 42, W252–W258 (2014).

    CAS  Article  Google Scholar 

  68. 68.

    Kelley, L. A., Mezulis, S., Yates, C. M., Wass, M. N. & Sternberg, M. J. The Phyre2 web portal for protein modeling, prediction and analysis. Nat. Protoc. 10, 845–858 (2015).

    CAS  Article  Google Scholar 

  69. 69.

    Emsley, P. & Cowtan, K. Coot: model-building tools for molecular graphics. Acta Crystallogr. D. Biol. Crystallogr. 60, 2126–2132 (2004).

    Article  Google Scholar 

  70. 70.

    Ding, Y. H. et al. Modeling protein excited-state structures from “over-length” chemical cross-links. J. Biol. Chem. 292, 1187–1196 (2017).

    CAS  Article  Google Scholar 

  71. 71.

    Herzog, F. et al. Structural probing of a protein phosphatase 2 A network by chemical cross-linking and mass spectrometry. Science 337, 1348–1352 (2012).

    CAS  Article  Google Scholar 

  72. 72.

    Erzberger, J. P. et al. Molecular architecture of the 40S·eIF1·eIF3 translation initiation complex. Cell 158, 1123–1135 (2014).

    CAS  Article  Google Scholar 

  73. 73.

    Adams, P. D. et al. PHENIX: a comprehensive Python-based system for macromolecular structure solution. Acta Crystallogr. D Biol. Crystallogr. 66, 213–221 (2010).

    Google Scholar 

  74. 74.

    Chen, V. B. et al. MolProbity: all-atom structure validation for macromolecular crystallography. Acta Crystallogr. D. Biol. Crystallogr. 66, 12–21 (2010).

    CAS  Article  Google Scholar 

Download references


We are grateful to J. Lei, Y. Xu, Z. Yan, Q. Zhou, J. Wang and T. Yang for their help with cryo-EM experiments, structure determination and model building. We thank the Beijing National Protein Science Facility at Tsinghua University and Electron Microscopy Resource Laboratory at the University of Pennsylvania for support. This work is supported by a National Institute of Health grant (R01 GM111128) to W.G. and grants from the National Science Foundation of China (Grant 31530018), Beijing Municipal Science & Technology Commission (Grant Z161100000116034) and National Key Research and Development Program of MOST (Grant 2016YFA0501100) to H.-W.W.

Author information




W.G. initiated the project. W.G. and H.-W.W. designed and supervised the experiments. K.M., G.L., P.Y., and Y.L. purified the exocyst complex. Y.L. and K.M. collected the EM data. Y.L., H.-W.W., J.W., K.M. and J.-J.L. analyzed the EM data and generated the EM map. G.S., Y.D. and M.-Q.D. performed the CXMS experiments. K.M., G.S., Y.L., M.-Q.D., W.G. and H.-W.W. analyzed the CXMS data. K.M., Y.L., H.-W.W. and W.G. built the atomic model. K.M., H.-W.W., W.G. and X.W. analyzed the structure. S.W. performed the microscopy of Sec3 mutations. S.W. and K.M. performed the secretion assays. K.M., S.W. and W.G. analyzed the cell biology data. W.G., K.M. and Y.L. wrote the draft. W.G., K.M., H.-W.W. and X.W. edited the manuscript.

Corresponding authors

Correspondence to Hong-Wei Wang or Wei Guo.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Integrated supplementary information

Supplementary Figure 1 Flow chart of data processing of the cryo-EM dataset.

Refer to the MATERIALS AND METHODS for details. The 3-D map in the red circle indicates a class with Sec3 in different conformation.

Supplementary Figure 2 The cryo-EM data processing results.

a. Cryo-EM 2-D class averages of the exocyst complexes purified from different strains with various subunit tagging. The red arrowheads indicate the density of C-terminally fused GFP in the corresponding complexes. CBD, calmodulin-binding domain. b. The full gallery of reference-free 2-D class averages that were included in the final 3-D reconstitution of the exocyst complex. The scale bar represents 20 nm. c. The angular distribution for the final 3-D reconstitution. The length of each cylinder is proportional to the number of particles for that view. d. The gold-standard FSC curve for the map refined with a mask around the body and distortion magnification correction. e. The Resmap of the 3-D reconstruction showing local resolutions.

Supplementary Figure 3 Flow chart and summary of the model building.

Regions built in each step are shown above the arrows, and modeling strategy is shown below the arrows. Back view maps are shown in the upper panel. Front view maps are shown in the lower panel. Also see Supplementary Video 1 for step-by-step exocyst model building and Supplementary Video 2 for CXMS analysis of the exocyst complex.

Supplementary Figure 4 Rigid docking and model building of Sec6, Sec10, Sec15, Exo70 and Exo84.

a-c. Rigid docking of Exo84 (a.a.525-753), Exo70 (a.a.67-623) and Sec6 (a.a.411-805), respectively. Exo70 (a.a. 67-623) was split into two domains for separate docking. Blue, Exo70 (a.a. 67-344); yellow, Exo70 (a.a. 345-623). The spheres in c indicate the Cα atoms of labeled lysine residues involved in the cross-linking. Distances between Cα-Cα of the crosslinked residues are shown above the dashed lines. d. Rigid docking of Sec10 based on Zebrafish Sec10(a.a.195-708) (PDB code: 5H11), Minimal adjustment was applied to fit the model into the map (Purple, original docking; cyan, docking with adjustment). e. Docking result of Exo84 PH domain based on rat Exo84(a.a.171-283) (PDB code: 1ZC4). f. Docking result of Sec15 C-terminal domain based on Drosophila Sec15(a.a.382-699) (PDB code: 2A2F). The map is set at high (left) and low threshold (right), respectively. g-k. Final models of Sec6, Sec10, Sec15, Exo70 and Exo84.

Supplementary Figure 5 De novo model building of Sec5, Sec8 and Sec3.

a-c. Model building of Sec5, Sec8 and Sec3. Close-up views show the cross-linking pairs that guided model building. Residue numbers are assigned based on secondary structure prediction. The spheres in each model indicate the Cα atoms of labeled residues in the cross-linking. Distances between Cα-Cα of the crosslinks are shown above the dashed lines (golden, Sec3; green, Sec5; purple, Sec6; forest green, Sec8; cyan, Sec10; blue, Exo70; pink, Exo84).

Supplementary Figure 6 Representative EM density maps for the yeast exocyst complex.

Several fragments of Sec5 (green), Sec15 (red) and Exo84 (pink) are shown, with the starting and ending residues labeled. Clear helical pitches can be seen in each of these fragments. While Sec5 and Sec15 are consisted of poly-alanine to trace the main chain, Exo84 (a.a. 525- 753) docked with crystal structure (PDB code: 2D2S) retains its side chains. Partial Exo84 side chains are displayed to show the consistency with the protuberances of the map.

Supplementary Figure 7 Secondary structure prediction of the exocyst subunits.

The diagram was generated with results from Predictprotein (brown, helix; blue, beta-strand). Amino acid numbers were shown in the scale bar above. The CorEx motifs of the exocyst subunits are indicated by black lines. The long linker region between the PH domain and the CorEx motif of Sec3 is indicated by the green line.

Figure Supplementary 8 Docking of Sec3 PH domain suggests the plasma membrane-binding surface of the exocyst complex.

The PH domain of Sec3 was docked to the rear bottom of the back layer based on its cross-linking with Sec15 (see text). The interface between the exocyst complex and the plasma membrane, as determined by the interactions of PI(4,5)P2 with Exo70 C-terminus and Sec3 PH domain (Yamashita, M. et al., Nat. Struct. Mol. Biol. 17, 180–186, 2010; He, B. et al., EMBO J. 26, 4053–4065, 2007), is modeled to the rear face of the back layer of the exocyst complex. The dash line connecting the PH domain and the built Sec3 model indicates the linker between the two regions. Golden, Sec3; blue, Exo70; red, Sec15. The red spheres mimic the phosphates of PI(4,5)P2.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–8, Supplementary Tables 1 and 2 and Supplementary Note 1

Life Sciences Reporting Summary

Supplementary Dataset 1

Results of mass spectrometric analysis of DSS/BS3 cross-linked intermolecular residues in the exocyst complex

Supplementary Dataset 2

Results of mass spectrometric analysis of DSS/BS3 cross-linked intramolecular residues in the exocyst complex

Supplementary Dataset 3

Results of mass spectrometric analysis of EDC cross-linked residues in the exocyst complex

Supplementary Dataset 4

Raw SDS-PAGE and western blot images

Supplementary Video 1

Model building of the exocyst complex. Step-by-step model building process is shown. First, available crystal structures from yeast were docked into the map. Then, structures predicted from homology structures were docked and mildly adjusted to fit the map. Next, de novo model extension was performed based on the map connectivity and secondary structure prediction. After that, the density for Sec5 and Sec8 was assigned with the guidance of the CXMS information and de novo model building was performed based on map connectivity and secondary structure prediction. Finally, the remaining density was assigned to Sec3 and its model was manually built as did with other subunits.

Supplementary Video 2

CXMS analysis of the exocyst complex. All confident (E value<1E–4 and counts>2) intermolecular and intramolecular residue pairs cross-linked by BS3, DSS and EDC are mapped onto the exocyst model. Crosslinks with Cα–Cα distance> 35 Å are displayed as outliers. In total, 280 crosslinks are shown, of which 11 are outliers.

Supplementary Video 3

The assembly of the exocyst complex. The hierarchical assembly of the exocyst complex is displayed. First, Sec3–Sec5, Sec6–Sec8, Sec10–Sec15 and Exo70–Exo84 formed heterodimer through their CorEx motifs. Then the four-helical bundle formation with the CorEx motifs of Sec3–Sec5–Sec6–Sec8 and Sec10–Sec15–Exo70–Exo84 results in the subcomplex 1 and subcomplex II. Finally, the holo-exocyst complex was assembled through the interactions between subcomplex I and subcomplex II.

Source data

Source Data, Figure 6c

Source data for Figure 6c

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Mei, K., Li, Y., Wang, S. et al. Cryo-EM structure of the exocyst complex. Nat Struct Mol Biol 25, 139–146 (2018).

Download citation

Further reading


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing