Skip to main content

Thank you for visiting nature.com. 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.

  • Article
  • Published:

Proximity-induced caspase-9 activation on a DNA origami-based synthetic apoptosome

Nature Catalysis volume 3, pages 295–306 (2020)Cite this article

Subjects

Abstract

Living cells regulate key cellular processes by spatial organization of catalytically active proteins in higher-order signalling complexes. These act as organizing centres to facilitate proximity-induced activation and inhibition of multiple intrinsically weakly associating signalling components, which makes elucidation of the underlying protein–protein interactions challenging. Here we show that DNA origami nanostructures provide a programmable molecular platform for the systematic analysis of signalling proteins by engineering a synthetic DNA origami-based version of the apoptosome, a multiprotein complex that regulates apoptosis by colocalizing multiple caspase-9 monomers. Tethering of both wild-type and inactive caspase-9 variants to a DNA origami platform demonstrates that enzymatic activity is induced by proximity-driven dimerization with half-of-sites reactivity and, furthermore, reveals a multivalent activity enhancement in oligomers of three and four enzymes. Our results offer fundamental insights in caspase-9 activity regulation and demonstrate that DNA origami-based protein assembly platforms have the potential to inform the function of other multi-enzyme complexes involved in inflammation, innate immunity and cell death.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: General concept and design elements for the construction of a DNA-based synthetic apoptosome.
Fig. 2: Characterization of caspase-9 assembly onto DNA origami nanostructures.
Fig. 3: Activation of caspase-9 occurs by distance-dependent dimerization of tethered monomers.
Fig. 4: Colocalization of more than two caspase-9 monomers leads to enhanced enzymatic activity.
Fig. 5: Enzymatic activity of the caspase-9 dimer originates from a single catalytic site.

Similar content being viewed by others

Data availability

The data that support the plots within this paper and other findings of this study are available from the corresponding author on reasonable request.

Code availability

Custom-written code for the computer models and simulations that support the experimental findings in this study is available from the corresponding author on reasonable request.

References

  1. Bhattacharyya, R. P., Reményi, A., Yeh, B. J. & Lim, W. A. Domains, motifs, and scaffolds: the role of modular interactions in the evolution and wiring of cell signaling circuits. Annu. Rev. Biochem. 75, 655–680 (2006).

    Article  CAS  PubMed  Google Scholar 

  2. Park, H. H. et al. The death domain superfamily in intracellular signaling of apoptosis and inflammation. Annu. Rev. Immunol. 25, 561–586 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Good, M. C., Zalatan, J. G. & Lim, W. A. Scaffold proteins: hubs for controlling the flow of cellular information. Science 332, 680–686 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Wu, H. Higher-order assemblies in a new paradigm of signal transduction. Cell 153, 287–292 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Kagan, J. C., Magupalli, V. G. & Wu, H. SMOCs: supramolecular organizing centres that control innate immunity. Nat. Rev. Immunol. 14, 821–826 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Lin, S.-C., Lo, Y.-C. & Wu, H. Helical assembly in the MyD88–IRAK4–IRAK2 complex in TLR/IL-1R signalling. Nature 465, 885–890 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Park, S.-H., Zarrinpar, A. & Lim, W. A. Rewiring MAP kinase pathways using alternative scaffold assembly mechanisms. Science 299, 1061–1064 (2003).

    Article  CAS  PubMed  Google Scholar 

  8. Tan, Y. & Kagan, J. C. Innate immune signaling organelles display natural and programmable signaling flexibility. Cell 177, 384–398.e11 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Kholodenko, B. N., Hancock, J. F. & Kolch, W. Signalling ballet in space and time. Nat. Rev. Mol. Cell Biol. 11, 414–426 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Castellana, M. et al. Enzyme clustering accelerates processing of intermediates through metabolic channeling. Nat. Biotechnol. 32, 1011–1018 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Lim, W. A. Designing customized cell signalling circuits. Nat. Rev. Mol. Cell Biol. 11, 393–403 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Conrado, R. J., Varner, J. D. & DeLisa, M. P. Engineering the spatial organization of metabolic enzymes: mimicking nature’s synergy. Curr. Opin. Biotechnol. 19, 492–499 (2008).

    Article  CAS  PubMed  Google Scholar 

  13. Idan, O. & Hess, H. Engineering enzymatic cascades on nanoscale scaffolds. Curr. Opin. Biotechnol. 24, 606–611 (2013).

    Article  CAS  PubMed  Google Scholar 

  14. Chen, Y.-J., Groves, B., Muscat, R. A. & Seelig, G. DNA nanotechnology from the test tube to the cell. Nat. Nanotechnol. 10, 748–760 (2015).

    Article  CAS  PubMed  Google Scholar 

  15. Seeman, N. C. & Sleiman, H. F. DNA nanotechnology. Nat. Rev. Mater. 3, 17068 (2017).

    Article  CAS  Google Scholar 

  16. Engelen, W., Janssen, B. M. G. & Merkx, M. DNA-based control of protein activity. Chem. Commun. 52, 3598–3610 (2016).

    Article  CAS  Google Scholar 

  17. Fu, J., Liu, M., Liu, Y. & Yan, H. Spatially-interactive biomolecular networks organized by nucleic acid nanostructures. Acc. Chem. Res. 45, 1215–1226 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Rothemund, P. W. K. Folding DNA to create nanoscale shapes and patterns. Nature 440, 297–302 (2006).

    Article  CAS  PubMed  Google Scholar 

  19. Voigt, N. V. et al. Single-molecule chemical reactions on DNA origami. Nat. Nanotechnol. 5, 200–203 (2010).

    Article  CAS  PubMed  Google Scholar 

  20. Saccà, B. & Niemeyer, C. M. Functionalization of DNA nanostructures with proteins. Chem. Soc. Rev. 40, 5910–5921 (2011).

    Article  CAS  PubMed  Google Scholar 

  21. Udomprasert, A. et al. Amyloid fibrils nucleated and organized by DNA origami constructions. Nat. Nanotechnol. 9, 537–541 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Xu, W. et al. A programmable DNA origami platform to organize SNAREs for membrane fusion. J. Am. Chem. Soc. 138, 4439–4447 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Funke, J. J. et al. Uncovering the forces between nucleosomes using DNA origami. Sci. Adv. 2, e1600974 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Le, J. V. et al. Probing nucleosome stability with a DNA origami nanocaliper. ACS Nano 10, 7073–7084 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Ketterer, P. et al. DNA origami scaffold for studying intrinsically disordered proteins of the nuclear pore complex. Nat. Commun. 9, 902 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Masubuchi, T. et al. Construction of integrated gene logic-chip. Nat. Nanotechnol. 13, 933–940 (2018).

    Article  CAS  PubMed  Google Scholar 

  27. Zhao, Z. et al. Nanocaged enzymes with enhanced catalytic activity and increased stability against protease digestion. Nat. Commun. 7, 10619 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Linko, V., Eerikäinen, M. & Kostiainen, M. A. A modular DNA origami-based enzyme cascade nanoreactor. Chem. Commun. 51, 5351–5354 (2015).

    Article  CAS  Google Scholar 

  29. Wilner, O. I. et al. Enzyme cascades activated on topologically programmed DNA scaffolds. Nat. Nanotechnol. 4, 249–254 (2009).

    Article  CAS  PubMed  Google Scholar 

  30. Fu, J., Liu, M., Liu, Y., Woodbury, N. W. & Yan, H. Interenzyme substrate diffusion for an enzyme cascade organized on spatially addressable DNA nanostructures. J. Am. Chem. Soc. 134, 5516–5519 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Timm, C. & Niemeyer, C. M. Assembly and purification of enzyme-functionalized DNA origami structures. Angew. Chem. Int Ed. 54, 6745–6750 (2015).

    Article  CAS  Google Scholar 

  32. Ngo, T. A., Nakata, E., Saimura, M. & Morii, T. Spatially organized enzymes drive cofactor-coupled cascade reactions. J. Am. Chem. Soc. 138, 3012–3021 (2016).

    Article  CAS  PubMed  Google Scholar 

  33. Fu, J. et al. Multi-enzyme complexes on DNA scaffolds capable of substrate channelling with an artificial swinging arm. Nat. Nanotechnol. 9, 531–536 (2014).

    Article  CAS  PubMed  Google Scholar 

  34. Riedl, S. J. & Salvesen, G. S. The apoptosome: signalling platform of cell death. Nat. Rev. Mol. Cell Biol. 8, 405–413 (2007).

    Article  CAS  PubMed  Google Scholar 

  35. Hu, Q. et al. Molecular determinants of caspase-9 activation by the Apaf-1 apoptosome. Proc. Natl Acad. Sci. USA 111, 16254–16261 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Li, Y. et al. Mechanistic insights into caspase-9 activation by the structure of the apoptosome holoenzyme. Proc. Natl Acad. Sci. USA 114, 1542–1547 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Cheng, T. C., Hong, C., Akey, I. V., Yuan, S. & Akey, C. W. A near atomic structure of the active human apoptosome. eLife 5, e17755 (2016).

    Article  PubMed  PubMed Central  Google Scholar 

  38. Renatus, M., Stennicke, H. R., Scott, F. L., Liddington, R. C. & Salvesen, G. S. Dimer formation drives the activation of the cell death protease caspase 9. Proc. Natl Acad. Sci. USA 98, 14250–14255 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Niemeyer, C. M. Semisynthetic DNA–protein conjugates for biosensing and nanofabrication. Angew. Chem. Int. Ed. 49, 1200–1216 (2010).

    Article  CAS  Google Scholar 

  40. Chin, J. W. et al. Addition of p-azido-l-phenylalanine to the genetic code of Escherichia coli. J. Am. Chem. Soc. 124, 9026–9027 (2002).

    Article  CAS  PubMed  Google Scholar 

  41. Chin, J. W. Expanding and reprogramming the genetic code of cells and animals. Annu. Rev. Biochem. 83, 379–408 (2014).

    Article  CAS  PubMed  Google Scholar 

  42. Liu, C. C. & Schultz, P. G. Adding new chemistries to the genetic code. Annu. Rev. Biochem. 79, 413–444 (2010).

    Article  CAS  PubMed  Google Scholar 

  43. Marth, G. et al. Precision templated bottom-up multiprotein nanoassembly through defined click chemistry linkage to DNA. ACS Nano 11, 5003–5010 (2017).

    Article  CAS  PubMed  Google Scholar 

  44. Glettenberg, M. & Niemeyer, C. M. Tuning of peroxidase activity by covalently tethered DNA oligonucleotides. Bioconjug. Chem. 20, 969–975 (2009).

    Article  CAS  PubMed  Google Scholar 

  45. Trads, J. B., Tørring, T. & Gothelf, K. V. Site-selective conjugation of native proteins with DNA. Acc. Chem. Res. 50, 1367–1374 (2017).

    Article  CAS  PubMed  Google Scholar 

  46. Sancho Oltra, N., Bos, J. & Roelfes, G. Control over enzymatic activity by DNA-directed split enzyme reassembly. ChemBioChem 11, 2255–2258 (2010).

    Article  CAS  Google Scholar 

  47. Yin, Q. et al. Caspase-9 holoenzyme is a specific and optimal procaspase-3 processing machine. Mol. Cell 22, 259–268 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Pop, C., Timmer, J., Sperandio, S. & Salvesen, G. S. The apoptosome activates caspase-9 by dimerization. Mol. Cell 22, 269–275 (2006).

    Article  CAS  PubMed  Google Scholar 

  49. Chao, Y. et al. Engineering a dimeric caspase-9: a re-evaluation of the induced proximity model for caspase activation. PLoS Biol. 3, e183 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Den Hamer, A. et al. Small-molecule-induced and cooperative enzyme assembly on a 14-3-3 scaffold. ChemBioChem 18, 331–335 (2017).

    Article  CAS  Google Scholar 

  51. Hu, Q., Wu, D., Chen, W., Yan, Z. & Shi, Y. Proteolytic processing of the caspase-9 zymogen is required for apoptosome-mediated activation of caspase-9. J. Biol. Chem. 288, 15142–15147 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Shiozaki, E. N., Chai, J. & Shi, Y. Oligomerization and activation of caspase-9, induced by Apaf-1 CARD. Proc. Natl Acad. Sci. USA 99, 4197–4202 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Wu, C.-C. et al. The Apaf-1 apoptosome induces formation of caspase-9 homo- and heterodimers with distinct activities. Nat. Commun. 7, 13565 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Huber, K. L., Serrano, B. P. & Hardy, J. A. Caspase-9 CARD: core domain interactions require a properly formed active site. Biochem. J. 475, 1177–1196 (2018).

    Article  CAS  PubMed  Google Scholar 

  55. Saccà, B. et al. Orthogonal protein decoration of DNA origami. Angew. Chem. Int Ed. 49, 9378–9383 (2010).

    Article  CAS  Google Scholar 

  56. Rosier, B. J. H. M. et al. Incorporation of native antibodies and Fc-fusion proteins on DNA nanostructures via a modular conjugation strategy. Chem. Commun. 53, 7393–7396 (2017).

    Article  CAS  Google Scholar 

  57. Baker, M. A. B. et al. Dimensions and global twist of single-layer DNA origami measured by small-angle X-ray scattering. ACS Nano 12, 5791–5799 (2018).

    Article  CAS  PubMed  Google Scholar 

  58. Snodin, B. E. K., Schreck, J. S., Romano, F., Louis, A. A. & Doye, J. P. K. Coarse-grained modelling of the structural properties of DNA origami. Nucleic Acids Res. 47, 1585–1597 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. Sa-Ardyen, P., Vologodskii, A. V. & Seeman, N. C. The flexibility of DNA double crossover molecules. Biophys. J. 84, 3829–3837 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  60. Zhang, Y., Tsitkov, S. & Hess, H. Proximity does not contribute to activity enhancement in the glucose oxidase–horseradish peroxidase cascade. Nat. Commun. 7, 13982 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  61. Stennicke, H. R. & Salvesen, G. S. Biochemical characteristics of caspases-3, -6, -7, and -8. J. Biol. Chem. 272, 25719–25723 (1997).

    Article  CAS  PubMed  Google Scholar 

  62. Wingard Jr., L. B., Katchalski-Katzir, E. & Goldstein, L. Applied Biochemistry and Bioengineering. Volume 1: Immobilized Enzyme Principles (Academic, 1976).

  63. Page, M. I. & Jencks, W. P. Entropic contributions to rate accelerations in enzymic and intramolecular reactions and the chelate effect. Proc. Natl Acad. Sci. USA 68, 1678–1683 (1971).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  64. Van Valen, D., Haataja, M. & Phillips, R. Biochemistry on a leash: the roles of tether length and geometry in signal integration proteins. Biophys. J. 96, 1275–1292 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  65. Salvesen, G. S. & Duckett, C. S. IAP proteins: blocking the road to death’s door. Nat. Rev. Mol. Cell Biol. 3, 401–410 (2002).

    Article  CAS  PubMed  Google Scholar 

  66. Shiozaki, E. N. et al. Mechanism of XIAP-mediated inhibition of caspase-9. Mol. Cell 11, 519–527 (2003).

    Article  CAS  PubMed  Google Scholar 

  67. Zaupa, G., Scrimin, P. & Prins, L. J. Origin of the dendritic effect in multivalent enzyme-like catalysts. J. Am. Chem. Soc. 130, 5699–5709 (2008).

    Article  CAS  PubMed  Google Scholar 

  68. Hill, T. L. & Levitzki, A. Subunit neighbor interactions in enzyme kinetics: half-of-the-sites reactivity in a dimer. Proc. Natl Acad. Sci. USA 77, 5741–5745 (1980).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  69. Biemann, H. P. & Koshland, D. E. Aspartate receptors of Escherichia coli and Salmonella typhimurium bind ligand with negative and half-of-the-sites cooperativity. Biochemistry 33, 629–634 (1994).

    Article  CAS  PubMed  Google Scholar 

  70. Vivoli, M., Pang, J. & Harmer, N. J. A half-site multimeric enzyme achieves its cooperativity without conformational changes. Sci. Rep. 7, 16529 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  71. Csizmok, V., Follis, A. V., Kriwacki, R. W. & Forman-Kay, J. D. Dynamic protein interaction networks and new structural paradigms in signaling. Chem. Rev. 116, 6424–6462 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  72. Levy, E. D., Erba, E. B., Robinson, C. V. & Teichmann, S. A. Assembly reflects evolution of protein complexes. Nature 453, 1262–1265 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  73. Bergendahl, L. T. & Marsh, J. A. Functional determinants of protein assembly into homomeric complexes. Sci. Rep. 7, 4932 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  74. Banani, S. F., Lee, H. O., Hyman, A. A. & Rosen, M. K. Biomolecular condensates: organizers of cellular biochemistry. Nat. Rev. Mol. Cell Biol. 18, 285–298 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  75. Shin, Y. & Brangwynne, C. P. Liquid phase condensation in cell physiology and disease. Science 357, eaaf4382 (2017).

    Article  CAS  PubMed  Google Scholar 

  76. Elbaz, J., Yin, P. & Voigt, C. A. Genetic encoding of DNA nanostructures and their self-assembly in living bacteria. Nat. Commun. 7, 11179 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  77. Praetorius, F. & Dietz, H. Self-assembly of genetically encoded DNA-protein hybrid nanoscale shapes. Science 355, eaam5488 (2017).

    Article  CAS  PubMed  Google Scholar 

  78. Delebecque, C. J., Lindner, A. B., Silver, P. A. & Aldaye, F. A. Organization of intracellular reactions with rationally designed RNA assemblies. Science 333, 470–474 (2011).

    Article  CAS  PubMed  Google Scholar 

  79. Weeks, S. D., Drinker, M. & Loll, P. J. Ligation independent cloning vectors for expression of SUMO fusions. Protein Expr. Purif. 53, 40–50 (2007).

    Article  CAS  PubMed  Google Scholar 

  80. Gasteiger, E. et al. in The Proteomics Protocols Handbook 571–607 (Humana Press, 2005).

  81. Tian, H., Sakmar, T. P. & Huber, T. A simple method for enhancing the bioorthogonality of cyclooctyne reagent. Chem. Commun. 52, 5451–5454 (2016).

    Article  CAS  Google Scholar 

  82. Den Hamer, A. et al. Bright bioluminescent BRET sensor proteins for measuring intracellular caspase activity. ACS Sens. 2, 729–734 (2017).

    Article  CAS  Google Scholar 

  83. Janssen, B. M. G., Engelen, W. & Merkx, M. DNA-directed control of enzyme–inhibitor complex formation: a modular approach to reversibly switch enzyme activity. ACS Synth. Biol. 4, 547–553 (2015).

    Article  CAS  PubMed  Google Scholar 

  84. Zadeh, J. N. et al. NUPACK: Analysis and design of nucleic acid systems. J. Comput. Chem. 32, 170–173 (2011).

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank J. van Dongen for help with the mass spectrometry analyses, N. van der Zon for initial protein expression experiments and G. Cremers for helpful discussions. The ICMS Animation Studio contributed the cartoons of DNA strands and the DNA origami structure. This work was supported by the European Research Council (project no. 677313 BioCircuit), an NWO-VIDI grant from the Netherlands Organisation for Scientific Research (723.016.003) and funding from the Ministry of Education, Culture and Science (Gravity programmes 024.001.035 and 024.003.013).

Author information

Authors and Affiliations

  1. Laboratory of Chemical Biology, Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, the Netherlands

    Bas J. H. M. Rosier, Anniek den Hamer, Luc Brunsveld & Tom F. A. de Greef

  2. Institute for Complex Molecular Systems, Eindhoven University of Technology, Eindhoven, the Netherlands

    Bas J. H. M. Rosier, Albert J. Markvoort, Berta Gumí Audenis, Job A. L. Roodhuizen, Anniek den Hamer, Luc Brunsveld & Tom F. A. de Greef

  3. Computational Biology Group, Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, the Netherlands

    Albert J. Markvoort, Job A. L. Roodhuizen & Tom F. A. de Greef

  4. Laboratory of Self-Organising Soft Matter and Laboratory of Macromolecular and Organic Chemistry, Department of Chemical Engineering and Chemistry, Eindhoven University of Technology, Eindhoven, the Netherlands

    Berta Gumí Audenis

  5. Institute for Molecules and Materials, Radboud University, Nijmegen, the Netherlands

    Tom F. A. de Greef

Authors
  1. Bas J. H. M. Rosier
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Albert J. Markvoort
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Berta Gumí Audenis
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Job A. L. Roodhuizen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Anniek den Hamer
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Luc Brunsveld
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Tom F. A. de Greef
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

B.J.H.M.R. designed the study, performed experiments, developed the geometric model, analysed the data, and wrote the manuscript. A.J.M. developed and derived the thermodynamic model and analysed the data. B.G.A. performed and analysed all AFM measurements. J.A.L.R. performed molecular dynamics simulations. A.d.H. performed initial protein expression and provided critical input for the experiments. L.B. supervised the study and provided critical feedback on the manuscript. T.F.A.d.G. conceived, designed and supervised the study, analysed the data and wrote the manuscript. All authors discussed the results and commented on the manuscript.

Corresponding authors

Correspondence to Luc Brunsveld or Tom F. A. de Greef.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

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

Supplementary information

Supplementary Information

Supplementary Methods, Figs. 1–41, Tables 1–7, Note 1 and references.

Reporting Summary

Supplementary Data 1

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Rosier, B.J.H.M., Markvoort, A.J., Gumí Audenis, B. et al. Proximity-induced caspase-9 activation on a DNA origami-based synthetic apoptosome. Nat Catal 3, 295–306 (2020). https://doi.org/10.1038/s41929-019-0403-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41929-019-0403-7

This article is cited by

Search

Advanced search

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