Using Single-Molecule Chemo-Mechanical Unfolding to Simultaneously Probe Multiple Structural Parameters in Protein Folding
Abstract
While single-molecule force spectroscopy has greatly advanced the study of protein folding, there are limitations to what can be learned from studying the effect of force alone. We developed a novel technique, chemo-mechanical unfolding, that combines multiple perturbants—force and chemical denaturant—to more fully characterize the folding process by simultaneously probing multiple structural parameters—the change in end-to-end distance, and solvent accessible surface area. Here, we describe the theoretical background, experimental design, and data analysis for chemo-mechanical unfolding experiments probing protein folding thermodynamics and kinetics. This technique has been applied to characterize parallel protein folding pathways, the protein denatured state, protein folding on the ribosome, and protein folding intermediates.
- Authors:
-
- DePauw Univ., Greencastle, IN (United States)
- Univ. of California, Berkeley, CA (United States); Chan Zuckerburg Biohub, San Francisco, CA (United States)
- Publication Date:
- Research Org.:
- Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC)
- OSTI Identifier:
- 1571996
- Grant/Contract Number:
- AC02-05CH11231
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Methods and Protocols
- Additional Journal Information:
- Journal Volume: 2; Journal Issue: 2; Journal ID: ISSN 2409-9279
- Publisher:
- MDPI
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; force spectroscopy; optical tweezers; chemo-mechanical unfolding; protein folding; denaturant; urea
Citation Formats
Guinn, Emily J., and Marqusee, Susan. Using Single-Molecule Chemo-Mechanical Unfolding to Simultaneously Probe Multiple Structural Parameters in Protein Folding. United States: N. p., 2019.
Web. doi:10.3390/mps2020032.
Guinn, Emily J., & Marqusee, Susan. Using Single-Molecule Chemo-Mechanical Unfolding to Simultaneously Probe Multiple Structural Parameters in Protein Folding. United States. https://doi.org/10.3390/mps2020032
Guinn, Emily J., and Marqusee, Susan. Sat .
"Using Single-Molecule Chemo-Mechanical Unfolding to Simultaneously Probe Multiple Structural Parameters in Protein Folding". United States. https://doi.org/10.3390/mps2020032. https://www.osti.gov/servlets/purl/1571996.
@article{osti_1571996,
title = {Using Single-Molecule Chemo-Mechanical Unfolding to Simultaneously Probe Multiple Structural Parameters in Protein Folding},
author = {Guinn, Emily J. and Marqusee, Susan},
abstractNote = {While single-molecule force spectroscopy has greatly advanced the study of protein folding, there are limitations to what can be learned from studying the effect of force alone. We developed a novel technique, chemo-mechanical unfolding, that combines multiple perturbants—force and chemical denaturant—to more fully characterize the folding process by simultaneously probing multiple structural parameters—the change in end-to-end distance, and solvent accessible surface area. Here, we describe the theoretical background, experimental design, and data analysis for chemo-mechanical unfolding experiments probing protein folding thermodynamics and kinetics. This technique has been applied to characterize parallel protein folding pathways, the protein denatured state, protein folding on the ribosome, and protein folding intermediates.},
doi = {10.3390/mps2020032},
journal = {Methods and Protocols},
number = 2,
volume = 2,
place = {United States},
year = {Sat Apr 20 00:00:00 EDT 2019},
month = {Sat Apr 20 00:00:00 EDT 2019}
}
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