Single molecule FRET investigation of pressure-driven unfolding of cold shock protein A

We demonstrate that fused silica capillaries are suitable for single molecule fluorescence resonance energy transfer (smFRET) measurements at high pressure with an optical quality comparable to the measurement on microscope coverslips. Therefore, we optimized the imaging conditions in a standard squ...

Full description

Saved in:
Bibliographic Details
Published in:The Journal of chemical physics 2018-03, Vol.148 (12), p.123336-123336
Main Authors: Schneider, Sven, Paulsen, Hauke, Reiter, Kim Colin, Hinze, Erik, Schiene-Fischer, Cordelia, Hübner, Christian G.
Format: Article
Language:English
Subjects:
Cold Shock Proteins and Peptides - chemistry
Fluorescence Resonance Energy Transfer
Pressure
Protein Unfolding
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:We demonstrate that fused silica capillaries are suitable for single molecule fluorescence resonance energy transfer (smFRET) measurements at high pressure with an optical quality comparable to the measurement on microscope coverslips. Therefore, we optimized the imaging conditions in a standard square fused silica capillary with an adapted arrangement and evaluated the performance by imaging the focal volume, fluorescence correlation spectroscopy benchmarks, and FRET measurements. We demonstrate single molecule FRET measurements of cold shock protein A unfolding at a pressure up to 2000 bars and show that the unfolded state exhibits an expansion almost independent of pressure.
ISSN:0021-9606
1089-7690
DOI:10.1063/1.5009662