Size-controlled assembly of phase separated protein condensates with interfacial protein cages

Hyeok Jin Oh, Yongsuk Lee, Haerang Hwang, Kibeom Hong, Hyeongjoo Choi, Jin Young Kang and Yongwon Jung ()
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Hyeok Jin Oh: Korea Advanced Institute of Science and Technology
Yongsuk Lee: Korea Advanced Institute of Science and Technology
Haerang Hwang: Korea Advanced Institute of Science and Technology
Kibeom Hong: Korea Advanced Institute of Science and Technology
Hyeongjoo Choi: Korea Advanced Institute of Science and Technology
Jin Young Kang: Korea Advanced Institute of Science and Technology
Yongwon Jung: Korea Advanced Institute of Science and Technology

Nature Communications, 2025, vol. 16, issue 1, 1-14

Abstract: Abstract Phase separation of specific proteins into liquid-like condensates is a key mechanism for forming membrane-less organelles, which organize diverse cellular processes in space and time. These protein condensates hold immense potential as biomaterials capable of containing specific sets of biomolecules with high densities and dynamic liquid properties. Despite their appeal, methods to manipulate protein condensate materials remain largely unexplored. Here, we present a one-pot assembly method to assemble coalescence-resistant protein condensates, ranging from a few μm to 100 nm in sizes, with surface-stabilizing protein cages. We discover that large protein cages (~30 nm), finely tuned to interact with condensates, efficiently localize on condensate surfaces and prevent the merging (coalescence) of condensates during phase separation. We precisely control condensate diameters by modulating condensate/cage ratios. In addition, the 3D structures of intact protein condensates with interfacial cages are visualized with cryo-electron tomography (ET). This work offers a versatile platform for designing size-controlled, surface-engineered protein condensate materials.

Date: 2025
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DOI: 10.1038/s41467-025-56391-y

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