Taylor-dispersion induced phase separation for efficient characterisation of protein condensate formation

Rasmus Krogh Norrild, Thomas Oliver Mason, Lars Boyens-Thiele, Soumik Ray, Joachim Berg Mortensen, Anatol W. Fritsch, Juan M. Iglesias-Artola, Louise Kjær Klausen, Emil Georg Palmkvist Stender, Henrik Jensen, Alexander Kai Buell

Research output: Contribution to journalJournal articleResearchpeer-review

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 nano-liters 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.
Original languageEnglish
Article numbere202404018
JournalANGEWANDTE CHEMIE
Volume63
Issue number25
Number of pages9
ISSN1433-7851
DOIs
Publication statusPublished - 2024

Keywords

  • Liqid-liquid phase separation
  • Biomolecular condensates
  • Microfluids
  • Protein solubility
  • Drug screening

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