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Force-Clamp Spectroscopy Monitors the Folding Trajectory of a Single Protein
Julio M. Fernandez* and
Hongbin Li
We used force-clamp atomic force micoscopy to measure the end-to-endlength of the small protein ubiquitin during its folding reactionat the single-molecule level. Ubiquitin was first unfolded andextended at a high force, then the stretching force was quenchedand protein folding was observed. The folding trajectories werecontinuous and marked by several distinct stages. The time takento fold was dependent on the contour length of the unfoldedprotein and the stretching force applied during folding. Thefolding collapse was marked by large fluctuations in the end-to-endlength of the protein, but these fluctuations vanished uponthe final folding contraction. These direct observations ofthe complete folding trajectory of a protein provide a benchmarktodetermine the physical basis of the folding reaction.
Department of Biological Sciences, Columbia University, New York, NY 10027, USA.
* To whom correspondence should be addressed. E-mail: jfernandez{at}columbia.edu
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