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Science 12 March 2004:
Vol. 303. no. 5664, pp. 1674 - 1678
DOI: 10.1126/science.1092497
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Reports

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-end length of the small protein ubiquitin during its folding reaction at the single-molecule level. Ubiquitin was first unfolded and extended at a high force, then the stretching force was quenched and protein folding was observed. The folding trajectories were continuous and marked by several distinct stages. The time taken to fold was dependent on the contour length of the unfolded protein and the stretching force applied during folding. The folding collapse was marked by large fluctuations in the end-to-end length of the protein, but these fluctuations vanished upon the final folding contraction. These direct observations of the complete folding trajectory of a protein provide a benchmarkto determine 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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