Controlling Drug Partitioning in Individual Protein Condensates through Laser-Induced Microscale Phase Transitions.

Autor:	Leppert A; Department of Cell and Molecular Biology, Uppsala University, S-75124 Uppsala, Sweden.; Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, S-17165 Solna, Sweden., Feng J; Bioproducts and Biosystems, Aalto University, Fi-00076 Aalto, Espoo, Finland., Railaite V; Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, S-17165 Solna, Sweden., Bohn Pessatti T; Department of Anatomy Physiology and Biochemistry, Swedish University of Agricultural Sciences, S-75007 Uppsala, Sweden., Cerrato CP; Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, S-17165 Solna, Sweden., Mörman C; Department of Biosciences and Nutrition, Karolinska Institutet, S-14157 Huddinge, Sweden.; Department of Biology and Chemistry, Paul Scherrer Institute, 5232 Villingen, Switzerland., Osterholz H; Department of Cell and Molecular Biology, Uppsala University, S-75124 Uppsala, Sweden., Lane DP; Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, S-17165 Solna, Sweden., Maia FRNC; Department of Cell and Molecular Biology, Uppsala University, S-75124 Uppsala, Sweden., Linder MB; Bioproducts and Biosystems, Aalto University, Fi-00076 Aalto, Espoo, Finland., Rising A; Department of Anatomy Physiology and Biochemistry, Swedish University of Agricultural Sciences, S-75007 Uppsala, Sweden.; Department of Biosciences and Nutrition, Karolinska Institutet, S-14157 Huddinge, Sweden., Landreh M; Department of Cell and Molecular Biology, Uppsala University, S-75124 Uppsala, Sweden.; Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, S-17165 Solna, Sweden.
Jazyk:	angličtina
Zdroj:	Journal of the American Chemical Society [J Am Chem Soc] 2024 Jul 17; Vol. 146 (28), pp. 19555-19565. Date of Electronic Publication: 2024 Jul 04.
DOI:	10.1021/jacs.4c06688
Abstrakt:	Gelation of protein condensates formed by liquid-liquid phase separation occurs in a wide range of biological contexts, from the assembly of biomaterials to the formation of fibrillar aggregates, and is therefore of interest for biomedical applications. Soluble-to-gel (sol-gel) transitions are controlled through macroscopic processes such as changes in temperature or buffer composition, resulting in bulk conversion of liquid droplets into microgels within minutes to hours. Using microscopy and mass spectrometry, we show that condensates of an engineered mini-spidroin (NT2repCT YF ) undergo a spontaneous sol-gel transition resulting in the loss of exchange of proteins between the soluble and the condensed phase. This feature enables us to specifically trap a silk-domain-tagged target protein in the spidroin microgels. Surprisingly, laser pulses trigger near-instant gelation. By loading the condensates with fluorescent dyes or drugs, we can control the wavelength at which gelation is triggered. Fluorescence microscopy reveals that laser-induced gelation significantly further increases the partitioning of the fluorescent molecules into the condensates. In summary, our findings demonstrate direct control of phase transitions in individual condensates, opening new avenues for functional and structural characterization.
Databáze:	MEDLINE
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