In vivo liquid–liquid phase separation protects amyloidogenic and aggregation-prone peptides during overexpression in Escherichia coli

Bartosz Gabryelczyk*, Reema Alag, Margaret Philips, Kimberly Low, Anandalakshmi Venkatraman, Bhuvaneswari Kannaian, Xiangyan Shi, Markus Linder, Konstantin Pervushin*, Ali Miserez*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)

Abstract

Studying pathogenic effects of amyloids requires homogeneous amyloidogenic peptide samples. Recombinant production of these peptides is challenging due to their susceptibility to aggregation and chemical modifications. Thus, chemical synthesis is primarily used to produce amyloidogenic peptides suitable for high-resolution structural studies. Here, we exploited the shielded environment of protein condensates formed via liquid–liquid phase separation (LLPS) as a protective mechanism against premature aggregation. We designed a fusion protein tag undergoing LLPS in Escherichia coli and linked it to highly amyloidogenic peptides, including β amyloids. We find that the fusion proteins form membraneless organelles during overexpression and remain fluidic-like. We also developed a facile purification method of functional Aβ peptides free of chromatography steps. The strategy exploiting LLPS can be applied to other amyloidogenic, hydrophobic, and repetitive peptides that are otherwise difficult to produce.

Original languageEnglish
Article numbere4292
JournalProtein Science
Volume31
Issue number5
DOIs
Publication statusPublished - May 2022
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2022 The Protein Society.

ASJC Scopus Subject Areas

  • Biochemistry
  • Molecular Biology

Keywords

  • amyloids
  • E. coli
  • liquid–liquid phase separation
  • membraneless organelles
  • protein condensates
  • protein tag
  • recombinant expression

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