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Science 29 August 2003:
Vol. 301. no. 5637, pp. 1233 - 1235
DOI: 10.1126/science.1085399
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Reports

Single-Molecule Measurement of Protein Folding Kinetics

Everett A. Lipman,1*{dagger} Benjamin Schuler,1,2* Olgica Bakajin,3 William A. Eaton1{ddagger}

In order to investigate the behavior of single molecules under conditions far from equilibrium, we have coupled a microfabricated laminar-flow mixer to a confocal optical system. This combination enables time-resolved measurement of Förster resonance energy transfer after an abrupt change in solution conditions. Observations of a small protein show the evolution of the intramolecular distance distribution as folding progresses. This technique can expose subpopulations, such as unfolded protein under conditions favoring the native structure, that would be obscured in equilibrium experiments.

1 Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, Building 5, Room 104, National Institutes of Health, Bethesda, MD 20892-0520, USA. 2 Physikalische Biochemie, Universität Potsdam, 14415 Potsdam, Germany. 3 Bio-security Nanosciences Laboratory, Chemistry and Materials Science, Lawrence Livermore National Laboratory, Livermore, CA 94550, USA.

* These authors contributedequally to this work.

{dagger} Present address: Department of Physics, University of California, Santa Barbara, CA 93106-9530, USA.

{ddagger} To whom correspondence should be addressed. E-mail: eaton{at}helix.nih.gov

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