DOI: 10.1126/science.1092497. Force-Clamp Spectroscopy Monitors the Folding Trajectory of a Single Protein

Force-Clamp Spectroscopy Monitors the Folding Trajectory of a Single Protein

10.1126/science.1092497

Crossref journal-article

American Association for the Advancement of Science (AAAS)

Science (221)

Abstract

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.

Bibliography

Fernandez, J. M., & Li, H. (2004). Force-Clamp Spectroscopy Monitors the Folding Trajectory of a Single Protein. Science, 303(5664), 1674â1678.

Authors 2

Julio M. Fernandez (first)
Hongbin Li (additional)

References 35 Referenced 494

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The use of a greatly improved piezoelectric actuator together with soft cantilevers makes it possible to control the force and the length of a single protein with pico-Newton and nanometer resolution. However it is still somewhat difficult to set the exact value of a low-force setting because our current force-clamp apparatus can be affected by small direct current offsets that affect our zero-force set point. These offsets do not affect our results at high stretching forces but can cause an unknown error when quenching the stretched protein to a low force. One way to independently measure the actual quenched force is to measure the magnitude of the elastic recoil that occurs immediately after relaxing the unfolded polymer chain to the lower force (e.g. steplike relaxation that coincides with downward arrows in Fig. 3). Given that in each case we know the contour length of the unfolded polypeptide we used the wormlike chain model of polymer elasticity to find the magnitude of the force quench for each elastic recoil event.
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We thank L. Li and K. Walther for helpful discussions and H. Huang for technical assistance. This work is funded by grants from NIH (J.M.F.).

Dates

Type	When
Created	21 years, 5 months ago (March 11, 2004, 5:06 p.m.)
Deposited	1 year, 7 months ago (Jan. 9, 2024, 11:09 p.m.)
Indexed	3 weeks, 2 days ago (Aug. 6, 2025, 9:38 a.m.)
Issued	21 years, 5 months ago (March 12, 2004)
Published	21 years, 5 months ago (March 12, 2004)
Published Print	21 years, 5 months ago (March 12, 2004)

Funders 0

None

BibTeX

@article{Fernandez_2004, title={Force-Clamp Spectroscopy Monitors the Folding Trajectory of a Single Protein}, volume={303}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.1092497}, DOI={10.1126/science.1092497}, number={5664}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Fernandez, Julio M. and Li, Hongbin}, year={2004}, month=mar, pages={1674–1678} }

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