Using Single-Molecule Chemo-Mechanical Unfolding to Simultaneously Probe Multiple Structural Parameters in Protein Folding

Guinn, Emily J.; Marqusee, Susan

doi:10.3390/mps2020032

Title: Using Single-Molecule Chemo-Mechanical Unfolding to Simultaneously Probe Multiple Structural Parameters in Protein Folding

Journal Article · Sat Apr 20 00:00:00 EDT 2019 · Methods and Protocols

DOI:https://doi.org/10.3390/mps2020032· OSTI ID:1571996

Guinn, Emily J. ^[1]; Marqusee, Susan ^[2]

DePauw Univ., Greencastle, IN (United States)
Univ. of California, Berkeley, CA (United States); Chan Zuckerburg Biohub, San Francisco, CA (United States)

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.

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Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1571996

Journal Information:: Methods and Protocols, Vol. 2, Issue 2; ISSN 2409-9279

Publisher:: MDPICopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
force spectroscopy
optical tweezers
chemo-mechanical unfolding
protein folding
denaturant
urea

Title: Using Single-Molecule Chemo-Mechanical Unfolding to Simultaneously Probe Multiple Structural Parameters in Protein Folding

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