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Constructing artificial gap junctions to mediate intercellular signal and mass transport

Nature Chemistry (2024)Cite this article

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Abstract

Natural gap junctions are a type of channel protein responsible for intercellular signalling and mass communication. However, the scope of applications for these proteins is limited as they cannot be prepared at a large scale and are unable to spontaneously insert into cell membranes in vitro. The construction of artificial gap junctions may provide an alternative strategy for preparing analogues of the natural proteins and bottom–up building blocks necessary for the synthesis of artificial cells. Here we show the construction of artificial gap junction channels from unimolecular tubular molecules consisting of alternately arranged positively and negatively charged pillar[5]arene motifs. These molecules feature a hydrophobic–hydrophilic–hydrophobic triblock structure that allows them to efficiently insert into two adjacent plasma membranes and stretch across the gap between the two membranes to form gap junctions. Similar to natural gap junction channels, the synthetic channels could mediate intercellular signal coupling and reactive oxygen species transmission, leading to cellular activity.

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Fig. 1: Structures of natural and artificial gap channels.
Fig. 2: Formation of tubular structures and their co-assembly with vesicles.
Fig. 3: Ion transport behaviours of channels 1–3.
Fig. 4: Ability of the channel to form artificial gap junctions.
Fig. 5: Intercellular signal coupling recording by whole-cell patch clamping (current-clamp mode) of two paired cells.
Fig. 6: Intercellular ROS transmission mediated by artificial gap junctions when C6 cells were incubated with FCCP-exposed cells.

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All data related to this study are available within the paper and its Supplementary Information. Source data are provided with this paper.

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Acknowledgements

We thank the National Key Research and Development Program of China (grant no. 2022YFA1203401, J.-L.H.), the National Natural Science Foundation of China (NSFC; grant nos. 21921003 and 21971046, J.-L.H.) and the Science and Technology Commission of Shanghai Municipality (STCSM; grant no. 22JC1403700, J.-L.H.) for financial support. We acknowledge X. Li (Shenzhen University) and G. Tang (Fudan University) for mass spectrum measurements and W. Wang (Fudan University) for helpful discussions on the MD simulations.

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Authors and Affiliations

  1. Department of Chemistry, Fudan University, Shanghai, China

    Yong-Hong Fu, Yi-Fei Hu, Tao Lin, Guo-Wei Zhuang, Ying-Lan Wang, Wen-Xue Chen, Zhan-Ting Li & Jun-Li Hou

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Contributions

Y.-H.F., Z.-T.L. and J.-L.H. conceived and designed the channel structures and experiments. Y.-H.F., Y.-F.H., T.L. and G.-W.Z. synthesized and characterized the channel molecules. Y.-H.F. investigated the self-assembly structure, and the intercellular signal and mass transport behaviours of the channel molecules. Y.-L.W. performed the MD simulations. W.-X.C. performed the 2D NMR experiments. Y.-H.F., Y.-F.H., Y.-L.W., W.-X.C. and J.-L.H. analysed the data and prepared the figures. Y.-H.F. and J.-L.H. wrote the paper.

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Correspondence to Jun-Li Hou.

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Nature Chemistry thanks Kazushi Kinbara, Chien-Lung Wang and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary Information

Supplementary Information

Experimental Procedures and Methods, Figs. 1–59 and Tables 1 and 2.

Reporting Summary

Supplementary Data 1

Initial and final configurations of the MD trajectories of channels 1–4.

Source data

Source Data Fig. 2

Source data for 2D 1H NMR NOESY spectrum (panel a) and DLS measurement plots (panel c).

Source Data Fig. 3

Source data for the current traces (panel a) and analysis on channel open events (panel b).

Source Data Fig. 4

Source data for panels a, b and c.

Source Data Fig. 5

Source data for the recorded membrane potential traces (panels b, c and d).

Source Data Fig. 6

Source data for panels b, c and d.

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Fu, YH., Hu, YF., Lin, T. et al. Constructing artificial gap junctions to mediate intercellular signal and mass transport. Nat. Chem. (2024). https://doi.org/10.1038/s41557-024-01519-8

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