Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Science 23 September 2005:
Vol. 309. no. 5743, pp. 2057 - 2060
DOI: 10.1126/science.1116702
Prev | Table of Contents | Next

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{dagger} Susan Marqusee1{dagger}

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 globule–like 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.

{dagger} To whom correspondence should be addressed. E-mail: carlos{at}alice.berkeley.edu; marqusee{at}berkeley.edu

Read the Full Text


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Fibronectin forms the most extensible biological fibers displaying switchable force-exposed cryptic binding sites.
E. Klotzsch, M. L. Smith, K. E. Kubow, S. Muntwyler, W. C. Little, F. Beyeler, D. Gourdon, B. J. Nelson, and V. Vogel (2009)
PNAS 106, 18267-18272
   Abstract »    Full Text »    PDF »
Single-molecule force spectroscopy distinguishes target binding modes of calmodulin.
J. P. Junker and M. Rief (2009)
PNAS 106, 14361-14366
   Abstract »    Full Text »    PDF »
Direct observation of an ensemble of stable collapsed states in the mechanical folding of ubiquitin.
S. Garcia-Manyes, L. Dougan, C. L. Badilla, J. Brujic, and J. M. Fernandez (2009)
PNAS 106, 10534-10539
   Abstract »    Full Text »    PDF »
Osmolyte-induced separation of the mechanical folding phases of ubiquitin.
S. Garcia-Manyes, L. Dougan, and J. M. Fernandez (2009)
PNAS 106, 10540-10545
   Abstract »    Full Text »    PDF »
Force Signaling in Biology.
J. Christof, M. Gebhardt, and M. Rief (2009)
Science 324, 1278-1280
   Abstract »    Full Text »    PDF »
Ligand-Dependent Equilibrium Fluctuations of Single Calmodulin Molecules.
J. P. Junker, F. Ziegler, and M. Rief (2009)
Science 323, 633-637
   Abstract »    Full Text »    PDF »
Stretching Single Talin Rod Molecules Activates Vinculin Binding.
A. del Rio, R. Perez-Jimenez, R. Liu, P. Roca-Cusachs, J. M. Fernandez, and M. P. Sheetz (2009)
Science 323, 638-641
   Abstract »    Full Text »    PDF »
Folding energy landscape and network dynamics of small globular proteins.
N. Hori, G. Chikenji, R. S. Berry, and S. Takada (2009)
PNAS 106, 73-78
   Abstract »    Full Text »    PDF »
Theory, analysis, and interpretation of single-molecule force spectroscopy experiments.
O. K. Dudko, G. Hummer, and A. Szabo (2008)
PNAS 105, 15755-15760
   Abstract »    Full Text »    PDF »
Optical tweezers for single cells.
H. Zhang and K.-K. Liu (2008)
J R Soc Interface 5, 671-690
   Abstract »    Full Text »    PDF »
Mechanical Biochemistry of Proteins One Molecule at a Time.
A. F. Oberhauser and M. Carrion-Vazquez (2008)
J. Biol. Chem. 283, 6617-6621
   Abstract »    Full Text »    PDF »
Solvent molecules bridge the mechanical unfolding transition state of a protein.
L. Dougan, G. Feng, H. Lu, and J. M. Fernandez (2008)
PNAS 105, 3185-3190
   Abstract »    Full Text »    PDF »
Single-molecule biophysics: at the interface of biology, physics and chemistry.
A. A Deniz, S. Mukhopadhyay, and E. A Lemke (2008)
J R Soc Interface 5, 15-45
   Abstract »    Full Text »    PDF »
Direct Observation of Chaperone-Induced Changes in a Protein Folding Pathway.
P. Bechtluft, R. G. H. van Leeuwen, M. Tyreman, D. Tomkiewicz, N. Nouwen, H. L. Tepper, A. J. M. Driessen, and S. J. Tans (2007)
Science 318, 1458-1461
   Abstract »    Full Text »    PDF »
Signatures of hydrophobic collapse in extended proteins captured with force spectroscopy.
K. A. Walther, F. Grater, L. Dougan, C. L. Badilla, B. J. Berne, and J. M. Fernandez (2007)
PNAS 104, 7916-7921
   Abstract »    Full Text »    PDF »
From the Cover: Single M13 bacteriophage tethering and stretching.
A. S. Khalil, J. M. Ferrer, R. R. Brau, S. T. Kottmann, C. J. Noren, M. J. Lang, and A. M. Belcher (2007)
PNAS 104, 4892-4897
   Abstract »    Full Text »    PDF »
Effect of force on mononucleosomal dynamics.
S. Mihardja, A. J. Spakowitz, Y. Zhang, and C. Bustamante (2006)
PNAS 103, 15871-15876
   Abstract »    Full Text »    PDF »
Hierarchic Finite Level Energy Landscape Model: TO DESCRIBE THE REFOLDING KINETICS OF PHOSPHOGLYCERATE KINASE.
S. Osvath, L. Herenyi, P. Zavodszky, J. Fidy, and G. Kohler (2006)
J. Biol. Chem. 281, 24375-24380
   Abstract »    Full Text »    PDF »
Anisotropic deformation response of single protein molecules.
H. Dietz, F. Berkemeier, M. Bertz, and M. Rief (2006)
PNAS 103, 12724-12728
   Abstract »    Full Text »    PDF »


To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)