A User's Guide for Phase Separation Assays with Purified Proteins

Publikation: Beitrag in FachzeitschriftForschungsartikelBeigetragenBegutachtung

Beitragende

  • Simon Alberti - , Professur für Zelluläre Biochemie, Max Planck Institute of Molecular Cell Biology and Genetics (Autor:in)
  • Shambaditya Saha - , Max Planck Institute of Molecular Cell Biology and Genetics (Autor:in)
  • Jeffrey B. Woodruff - , Max Planck Institute of Molecular Cell Biology and Genetics (Autor:in)
  • Titus M. Franzmann - , Professur für Zelluläre Biochemie, Max Planck Institute of Molecular Cell Biology and Genetics (Autor:in)
  • Jie Wang - , Max Planck Institute of Molecular Cell Biology and Genetics (Autor:in)
  • Anthony A. Hyman - , Max Planck Institute of Molecular Cell Biology and Genetics (Autor:in)

Abstract

The formation of membrane-less organelles and compartments by protein phase separation is an important way in which cells organize their cytoplasm and nucleoplasm. In vitro phase separation assays with purified proteins have become the standard way to investigate proteins that form membrane-less compartments. By now, various proteins have been purified and tested for their ability to phase separate and form liquid condensates in vitro. However, phase-separating proteins are often aggregation-prone and difficult to purify and handle. As a consequence, the results from phase separation assays often differ between labs and are not easily reproduced. Thus, there is an urgent need for high-quality proteins, standardized procedures, and generally agreed-upon practices for protein purification and conducting phase separation assays. This paper provides protocols for protein purification and guides the user through the practicalities of in vitro protein phase separation assays, including best-practice approaches and pitfalls to avoid. We believe that this compendium of protocols and practices will provide a useful resource for scientists studying the phase behavior of proteins.

Details

OriginalspracheEnglisch
Seiten (von - bis)4806-4820
Seitenumfang15
FachzeitschriftJournal of Molecular Biology
Jahrgang430
Ausgabenummer23
PublikationsstatusVeröffentlicht - 2 Nov. 2018
Peer-Review-StatusJa

Externe IDs

PubMed 29944854
ORCID /0000-0003-4017-6505/work/142253856

Schlagworte

Schlagwörter

  • low-complexity proteins, membrane-less compartment, membrane-less organelle, phase separation, prion-like protein, protein purification, Sup35

Bibliotheksschlagworte