Hsin-Liang Chen and Jack C.-C. Hsu contributed equally to this work.
Studying submicrosecond protein folding kinetics using a photolabile caging strategy and time-resolved photoacoustic calorimetry
Article first published online: 23 JUL 2010
Copyright © 2010 Wiley-Liss, Inc.
Proteins: Structure, Function, and Bioinformatics
Volume 78, Issue 14, pages 2973–2983, 1 November 2010
How to Cite
Chen, H.-L., Hsu, J. C.-C., Viet, M. H., Li, M. S., Hu, C.-K., Liu, C.-H., Luh, F. Y., Chen, S. S.-W., Chang, E. S.-H., Wang, A. H.-J., Hsu, M.-F., Fann, W. and Chen, R. P.-Y. (2010), Studying submicrosecond protein folding kinetics using a photolabile caging strategy and time-resolved photoacoustic calorimetry. Proteins, 78: 2973–2983. doi: 10.1002/prot.22823
- Issue published online: 8 SEP 2010
- Article first published online: 23 JUL 2010
- Manuscript Accepted: 2 JUL 2010
- Manuscript Revised: 24 JUN 2010
- Manuscript Received: 7 MAY 2010
- National Science Council, Taiwan, R.O.C.. Grant Numbers: NSC-95-2113-M-001-049, NSC-97-2113-M-001-014-MY2, NSC-96-2911-M-001-003-MY3, NSC-98-2911-I-001-028
- Academia Sinica. Grant Numbers: AS-94-TP-A01, AS-95-TP-A07
- Ministry of Science and Informatics in Poland. Grant Number: 202-204-234
- National Center for Theoretical Sciences in Taiwan
- protein folding;
- photoacoustic calorimetry
Kinetic measurement of protein folding is limited by the method used to trigger folding. Traditional methods, such as stopped flow, have a long mixing dead time and cannot be used to monitor fast folding processes. Here, we report a compound, 4-(bromomethyl)-6,7-dimethoxycoumarin, that can be used as a “photolabile cage” to study the early stages of protein folding. The folding process of a protein, RD1, including kinetics, enthalpy, and volume change, was studied by the combined use of a phototriggered caging strategy and time-resolved photoacoustic calorimetry. The cage caused unfolding of the photolabile protein, and then a pulse UV laser (∼10−9 s) was used to break the cage, leaving the protein free to refold and allowing the resolving of two folding events on a nanosecond time scale. This strategy is especially good for monitoring fast folding proteins that cannot be studied by traditional methods. Proteins 2010. © 2010 Wiley-Liss, Inc.