Pressure- and heat-induced protein unfolding in bacterial cells: crowding vs. sticking

Timothy Chen, Kapil Dave, Martin H W Gruebele

Research output: Contribution to journalLetter

Abstract

In-cell protein stability is increased by crowding, but can be reduced by destabilizing surface interactions. Will different denaturation techniques yield similar trends? Here, we apply pressure and thermal denaturation to green fluorescent protein/ReAsH-labeled yeast phosphoglycerate kinase (PGK) in Escherichia coli cells. Pressure denaturation is more two state-like in E. coli than in vitro, stabilizing the native state. Thermal denaturation destabilizes PGK in E. coli, unlike in mammalian cells. Results in wild-type MG1655 strain are corroborated in pressure-resistant J1 strain, where PGK is less prone to aggregation. Thus, destabilizing surface interactions overcome stabilizing crowding in the E. coli cytoplasm under thermal denaturation, but not under pressure denaturation.

Original languageEnglish (US)
Pages (from-to)1357-1365
Number of pages9
JournalFEBS Letters
Volume592
Issue number8
DOIs
StatePublished - Apr 2018

Fingerprint

Protein Unfolding
Denaturation
Phosphoglycerate Kinase
Hot Temperature
Escherichia coli
Pressure
Proteins
Protein Stability
Green Fluorescent Proteins
Cytoplasm
Yeasts
Yeast
Agglomeration
Cells

Keywords

  • E. coli
  • in-cell folding
  • pressure denaturation
  • quinary interaction

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Pressure- and heat-induced protein unfolding in bacterial cells : crowding vs. sticking. / Chen, Timothy; Dave, Kapil; Gruebele, Martin H W.

In: FEBS Letters, Vol. 592, No. 8, 04.2018, p. 1357-1365.

Research output: Contribution to journalLetter

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