DOI: 10.1073/pnas.96.7.3694. Mechanical and chemical unfolding of a single protein: A comparison

Mechanical and chemical unfolding of a single protein: A comparison

10.1073/pnas.96.7.3694

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

Is the mechanical unraveling of protein domains by atomic force microscopy (AFM) just a technological feat or a true measurement of their unfolding? By engineering a protein made of tandem repeats of identical Ig modules, we were able to get explicit AFM data on the unfolding rate of a single protein domain that can be accurately extrapolated to zero force. We compare this with chemical unfolding rates for untethered modules extrapolated to 0 M denaturant. The unfolding rates obtained by the two methods are the same. Furthermore, the transition state for unfolding appears at the same position on the folding pathway when assessed by either method. These results indicate that mechanical unfolding of a single protein by AFM does indeed reflect the same event that is observed in traditional unfolding experiments. The way is now open for the extensive use of AFM to measure folding reactions at the single-molecule level. Single-molecule AFM recordings have the added advantage that they define the reaction coordinate and expose rare unfolding events that cannot be observed in the absence of chemical denaturants.

Authors 7

Mariano Carrion-Vazquez (first)
Andres F. Oberhauser (additional)
Susan B. Fowler (additional)
Piotr E. Marszalek (additional)
Sheldon E. Broedel (additional)
Jane Clarke (additional)
Julio M. Fernandez (additional)

References 31 Referenced 920

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Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 10:39 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 5:31 p.m.)
Indexed	2 weeks, 3 days ago (Aug. 12, 2025, 5:37 p.m.)
Issued	26 years, 5 months ago (March 30, 1999)
Published	26 years, 5 months ago (March 30, 1999)
Published Online	26 years, 5 months ago (March 30, 1999)
Published Print	26 years, 5 months ago (March 30, 1999)

Funders 0

None

BibTeX

@article{Carrion_Vazquez_1999, title={Mechanical and chemical unfolding of a single protein: A comparison}, volume={96}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.96.7.3694}, DOI={10.1073/pnas.96.7.3694}, number={7}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Carrion-Vazquez, Mariano and Oberhauser, Andres F. and Fowler, Susan B. and Marszalek, Piotr E. and Broedel, Sheldon E. and Clarke, Jane and Fernandez, Julio M.}, year={1999}, month=mar, pages={3694–3699} }

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