Using Single-Molecule Chemo-Mechanical Unfolding to Simultaneously Probe Multiple Structural Parameters in Protein Folding
| Autor: | Emily J Guinn, Susan Marqusee |
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| Rok vydání: | 2019 |
| Předmět: |
1.1 Normal biological development and functioning
Kinetics chemo-mechanical unfolding Bioengineering force spectroscopy urea 010402 general chemistry 01 natural sciences Biochemistry Genetics and Molecular Biology (miscellaneous) Accessible surface area 03 medical and health sciences Structural Biology Underpinning research protein folding Protocol Molecule lcsh:QH301-705.5 030304 developmental biology 0303 health sciences Chemo mechanical Chemistry optical tweezers Force spectroscopy 0104 chemical sciences Folding (chemistry) denaturant Optical tweezers lcsh:Biology (General) Biophysics Protein folding Generic health relevance Biotechnology |
| Zdroj: | Methods and protocols, vol 2, iss 2 Methods and Protocols, Vol 2, Iss 2, p 32 (2019) Methods and Protocols |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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