Chapter title |
Single-Molecule Protein Folding Experiments Using High-Precision Optical Tweezers
|
---|---|
Chapter number | 14 |
Book title |
Optical Tweezers
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6421-5_14 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6419-2, 978-1-4939-6421-5
|
Authors |
Junyi Jiao, Aleksander A. Rebane, Lu Ma, Yongli Zhang, Jiao, Junyi, Rebane, Aleksander A., Ma, Lu, Zhang, Yongli |
Abstract |
How proteins fold from linear chains of amino acids to delicate three-dimensional structures remains a fundamental biological problem. Single-molecule manipulation based on high-resolution optical tweezers (OT) provides a powerful approach to study protein folding with unprecedented spatiotemporal resolution. In this method, a single protein or protein complex is tethered between two beads confined in optical traps and pulled. Protein unfolding induced by the mechanical force is counteracted by the spontaneous folding of the protein, reaching a dynamic equilibrium at a characteristic force and rate. The transition is monitored by the accompanying extension change of the protein and used to derive conformations and energies of folding intermediates and their associated transition kinetics. Here, we provide general strategies and detailed protocols to study folding of proteins and protein complexes using optical tweezers, including sample preparation, DNA-protein conjugation and methods of data analysis to extract folding energies and rates from the single-molecule measurements. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
United States | 1 | 100% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Scientists | 1 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 43 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 10 | 23% |
Student > Bachelor | 6 | 14% |
Student > Master | 4 | 9% |
Unspecified | 3 | 7% |
Researcher | 3 | 7% |
Other | 2 | 5% |
Unknown | 15 | 35% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 12 | 28% |
Physics and Astronomy | 5 | 12% |
Unspecified | 3 | 7% |
Agricultural and Biological Sciences | 2 | 5% |
Engineering | 2 | 5% |
Other | 3 | 7% |
Unknown | 16 | 37% |