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Superstructural ordering in self-sorting coacervate-based protocell networks

Nature Chemistry volume 16, pages 158–167 (2024)Cite this article

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Abstract

Bottom-up assembly of higher-order cytomimetic systems capable of coordinated physical behaviours, collective chemical signalling and spatially integrated processing is a key challenge in the study of artificial multicellularity. Here we develop an interactive binary population of coacervate microdroplets that spontaneously self-sort into chain-like protocell networks with an alternating sequence of structurally and compositionally dissimilar microdomains with hemispherical contact points. The protocell superstructures exhibit macromolecular self-sorting, spatially localized enzyme/ribozyme biocatalysis and interdroplet molecular translocation. They are capable of topographical reconfiguration using chemical or light-mediated stimuli and can be used as a micro-extraction system for macroscale biomolecular sorting. Our methodology opens a pathway towards the self-assembly of multicomponent protocell networks based on selective processes of coacervate droplet–droplet adhesion and fusion, and provides a step towards the spontaneous orchestration of protocell models into artificial tissues and colonies with ordered architectures and collective functions.

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Fig. 1: Spontaneous assembly, spatial segregation, self-organization and macroscopic sorting of coacervate-based protocell networks.
Fig. 2: Protocell network components.
Fig. 3: Protocell network assembly.
Fig. 4: Biomolecular organization and processing in protocell networks.
Fig. 5: Stimuli-induced reconfiguration in protocell networks.
Fig. 6: Protocell-network-mediated biomolecular extraction and macroscale sorting.

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Data availability

All data supporting the findings of this study are available within the article and its Supplementary Information, and from the corresponding authors on reasonable request. Source Data are provided with this paper.

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Acknowledgements

This work was supported by the Strategic Priority Research Program of the Chinese Academy of Sciences (grant no. XDB0480000) to Y.Q., the National Natural Science Foundation of China (grant nos. 22272183 and 22072159 to Y.Q., and 22172007 to Y.L.), the Science Fund for Creative Research Groups of the National Natural Science Foundation of China (grant no. 52221006) to Y.L., and the Fundamental Research Funds for the Central Universities (grant nos. buctrc202015 and PT2208 to Y.L). S.M. was funded by the ERC Advanced Grant Scheme (grant no. EC-2016-674 ADG 740235).

Author information

Author notes

  1. These authors contributed equally: Wenjing Mu, Liyan Jia.

Authors and Affiliations

  1. Beijing National Laboratory for Molecular Sciences (BNLMS), Laboratory of Polymer Physics and Chemistry, CAS Research/Education Center for Excellence in Molecular Sciences, Institute of Chemistry, Chinese Academy of Sciences, Beijing, China

    Wenjing Mu, Liyan Jia & Yan Qiao

  2. University of Chinese Academy of Sciences, Beijing, China

    Wenjing Mu, Liyan Jia & Yan Qiao

  3. Department of Chemical and Environmental Engineering, University of California, Riverside, CA, USA

    Musen Zhou & Jianzhong Wu

  4. State Key Laboratory of Chemical Resource Engineering, Beijing Laboratory of Biomedical Materials, Beijing University of Chemical Technology, Beijing, China

    Yiyang Lin

  5. Centre for Protolife Research and Centre for Organized Matter Chemistry, School of Chemistry, University of Bristol, Bristol, UK

    Stephen Mann

  6. Max Planck-Bristol Centre for Minimal Biology, School of Chemistry, University of Bristol, Bristol, UK

    Stephen Mann

  7. School of Materials Science and Engineering, Shanghai Jiao Tong University, Shanghai, P. R. China

    Stephen Mann

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  1. Wenjing Mu
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Contributions

W.M. and L.J. performed the experiments and analysed the data. M.Z. and J.W. performed computational experiments. Y.Q. led the project. Y.L., S.M. and Y.Q. conceived, designed and supervised the study, analysed the data and wrote the manuscript. All authors discussed the results and commented on the manuscript.

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Correspondence to Yiyang Lin, Stephen Mann or Yan Qiao.

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Supplementary information

Supplementary Information

Supplementary Video captions 1–4, materials and methods, Figs. 1–33, Tables 1–3 and references.

Supplementary Video 1

CLSM video showing the assembly of chain-like protocell networks comprising an alternating interconnected sequence of Nile-red-loaded DDAB/trans-AzoAsp2 (red fluorescence) and HPTS-loaded PDDA/trans-AzoAsp2 (green fluorescence) coacervate droplets are observed. Video is shown at ×540 of real-time speed at 3 fps. Total duration of recording in real time is 60 min; scale bar, 13 μm.

Supplementary Video 2

CLSM video showing HRP/H2O2-mediated peroxidation in coacervate-based protocell networks. Resorufin (red fluorescence) is produced and retained specifically within the DDAB/trans-AzoAsp2 domains. Video is shown at ×100 of real-time speed at 5 fps. Total duration of recording in real time is 600 s; scale bar, 5 μm.

Supplementary Video 3

CLSM video showing transfer of TAMRA-ssDNA from a DDAB/trans-AzoAsp2 droplet (red fluorescence at t = 0) to an adjacent PDDA/trans-AzoAsp2 domain (no fluorescence at t = 0) located in the same protocell chain. Video is shown at ×5 of real-time speed at 10 fps. Total duration of recording in real time is 38.5 s; scale bar, 10 μm.

Supplementary Video 4

Optical microscopy video showing UV-induced reconfiguration of protocell networks due to light-mediated trans-to-cis isomerization of azobenzene in ordered chains of DDAB/trans-AzoAsp2 and PDDA/trans-AzoAsp2 droplets. UV source; 325 nm < λ < 375 nm, 120 W short-arc Hg light. Video is shown at ×75 of real-time speed at 5 fps. Total duration of recording in real time is 10 min; scale bar, 5 μm.

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Mu, W., Jia, L., Zhou, M. et al. Superstructural ordering in self-sorting coacervate-based protocell networks. Nat. Chem. 16, 158–167 (2024). https://doi.org/10.1038/s41557-023-01356-1

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