Journal
CHEMPHYSCHEM
Volume 19, Issue 18, Pages 2290-2294Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cphc.201800534
Keywords
FRET; laser-induced temperature-jump; macromolecular crowding; protein folding; quinary interactions
Funding
- National Science Foundation (NSF) [NSF MCB 1413256, NSF MCB 1803786]
- PFC: Center for the Physics of Living Cells - NSF [PHY 1430124]
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Although biomolecules evolved to function in the cell, most biochemical assays are carried out invitro. In-cell studies highlight how steric and non-steric interactions modulate protein folding and interactions. VlsE and PGK present two extremes of chemical behavior in the cell: the extracellular protein VlsE is destabilized in eukaryotic cells, whereas the cytoplasmic protein PGK is stabilized. VlsE and PGK are benchmarks in a systematic series of solvation environments to distinguish contributions from non-steric and steric interactions to protein stability, compactness, and folding rate by comparing cell lysate, a crowding agent, ionic buffer and lysate buffer with in-cell results. As anticipated, crowding stabilizes proteins, causes compaction, and can speed folding. Protein flexibility determines its sensitivity to steric interactions or crowding. Non-steric interactions alone predict in-cell stability trends, while crowding provides an offset towards greater stabilization. We suggest that a simple combination of lysis buffer and Ficoll is an effective new invitro mimic of the intracellular environment on protein folding and stability.
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