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Direct Observation of the Three-State Folding of a Single Protein Molecule
Open accessAbstract
We used force-measuring optical tweezers to induce complete mechanical unfolding and refolding of individual Escherichia coli ribonuclease H (RNase H) molecules. The protein unfolds in a two-state manner and refolds through an intermediate that correlates with the transient molten globule–like intermediate observed in bulk studies. This intermediate displays unusual mechanical compliance and unfolds at substantially lower forces than the native state. In a narrow range of forces, the molecule hops between the unfolded and intermediate states in real time. Occasionally, hopping was observed to stop as the molecule crossed the folding barrier directly from the intermediate, demonstrating that the intermediate is on-pathway. These studies allow us to map the energy landscape of RNase H.
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Cited by
Stretching Single Talin Rod Molecules Activates Vinculin Binding
del Rio, A., Perez-Jimenez, R., Liu, R., Roca-Cusachs, P., Fernandez, J. M., & Sheetz, M. P. (2009) · Science · doi:10.1126/science.1162912 · Cited by 1,506
Dill, K. A., Ozkan, S. B., Shell, M. S., & Weikl, T. R. (2008) · Annual Review of Biophysics · doi:10.1146/annurev.biophys.37.092707.153558 · Cited by 1,163
Recent Advances in Optical Tweezers
Moffitt, J. R., Chemla, Y. R., Smith, S. B., & Bustamante, C. (2008) · Annual Review of Biochemistry · doi:10.1146/annurev.biochem.77.043007.090225 · Cited by 1,084
Self-citationOptical tweezers for single cells
Zhang, H., & Liu, K.-K. (2008) · Journal of The Royal Society Interface · doi:10.1098/rsif.2008.0052 · Cited by 768
Optical tweezers in single-molecule biophysics
Bustamante, C. J., Chemla, Y. R., Liu, S., & Wang, M. D. (2021) · Nature Reviews Methods Primers · doi:10.1038/s43586-021-00021-6 · Cited by 615
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Metadata
- Authors
- Cecconi, C., Shank, E. A., Bustamante, C., & Marqusee, S.
- Venue
- Science
- Published
- 2005-09-23
- Volume
- 309
- Issue
- 5743
- Pages
- 2057-2060
- ISSN
- 0036-8075 (ISSN-L)
- 1095-9203
- OpenAlex
- W2063616898
- Cited by
- 635
- References
- 0
- FWCI
- 12.45
- Type
- journal article
- Language
- en
- Open access
- green
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- Cecconi, C., Shank, E. A., Bustamante, C., & Marqusee, S. (2005). Direct Observation of the Three-State Folding of a Single Protein Molecule. Science. https://doi.org/10.1126/science.1116702
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