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- Title
Taylor Dispersion‐Induced Phase Separation for the Efficient Characterisation of Protein Condensate Formation.
- Authors
Norrild, Rasmus K.; Mason, Thomas O.; Boyens‐Thiele, Lars; Ray, Soumik; Mortensen, Joachim B.; Fritsch, Anatol W.; Iglesias‐Artola, Juan M.; Klausen, Louise K.; Stender, Emil G. P.; Jensen, Henrik; Buell, Alexander K.
- Abstract
Biomolecular condensates have emerged as important structures in cellular function and disease, and are thought to form through liquid‐liquid phase separation (LLPS). Thorough and efficient in vitro experiments are therefore needed to elucidate the driving forces of protein LLPS and the possibility to modulate it with drugs. Here we present Taylor dispersion‐induced phase separation (TDIPS), a method to robustly measure condensation phenomena using a commercially available microfluidic platform. It uses only nanoliters of sample, does not require extrinsic fluorescent labels, and is straightforward to implement. We demonstrate TDIPS by screening the phase behaviour of two proteins that form biomolecular condensates in vivo, PGL‐3 and Ddx4. Uniquely accessible to this method, we find an unexpected re‐entrant behaviour at very low ionic strength, where LLPS is inhibited for both proteins. TDIPS can also probe the reversibility of assemblies, which was shown for both α‐synuclein and for lysozyme, relevant for health and biotechnology, respectively. Finally, we highlight how effective inhibition concentrations and partitioning of LLPS‐modifying compounds can be screened highly efficiently.
- Subjects
PHASE separation; IONIC strength; ALPHA-synuclein; PROTEINS; CELL anatomy; CELL physiology
- Publication
Angewandte Chemie, 2024, Vol 136, Issue 25, p1
- ISSN
0044-8249
- Publication type
Article
- DOI
10.1002/ange.202404018