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Science 23 September 2005: Vol. 309. no. 5743, pp. 2057 - 2060 DOI: 10.1126/science.1116702
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Reports
Direct Observation of the Three-State Folding of a Single Protein Molecule
Ciro Cecconi,1,2*
Elizabeth A. Shank,1*
Carlos Bustamante,1,2,3
Susan Marqusee1
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 globulelike 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.
1 Department of Molecular and Cell Biology and Institute for Quantitative Biology
2 Department of Physics, Howard Hughes Medical Institute, University of California, Berkeley, CA 94720, USA.
3 Department of Physics, Howard Hughes Medical Institute, University of California, Berkeley, CA 94720, USA.
* These authors contributed equally to this work.
To whom correspondence should be addressed. E-mail: carlos{at}alice.berkeley.edu; marqusee{at}berkeley.edu
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