Research Article

Studying submicrosecond protein folding kinetics using a photolabile caging strategy and time-resolved photoacoustic calorimetry

  1. Hsin-Liang Chen1,2,†,
  2. Jack C.-C. Hsu1,3,†,
  3. Man Hoang Viet4,
  4. Mai Suan Li4,
  5. Chin-Kun Hu5,6,
  6. Chia-Hsun Liu1,
  7. Frederick Y. Luh1,
  8. Silvia S.-W. Chen1,
  9. Evan S.-H. Chang1,
  10. Andrew H.-J. Wang1,3,
  11. Min-Feng Hsu1,7,
  12. Wunshain Fann2,
  13. Rita P.-Y. Chen1,3,*

Article first published online: 23 JUL 2010

DOI: 10.1002/prot.22823

Issue

Proteins: Structure, Function, and Bioinformatics

Proteins: Structure, Function, and Bioinformatics

Volume 78, Issue 14, pages 2973–2983, 1 November 2010

Additional Information

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

Author Information

  1. 1

    Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan

  2. 2

    Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei 10617, Taiwan

  3. 3

    Institute of Biochemical Sciences, National Taiwan University, Taipei 10617, Taiwan

  4. 4

    Institute of Physics, Polish Academy of Sciences, 02-668 Warsaw, Poland

  5. 5

    Institute of Physics, Academia Sinica, Taipei 11529, Taiwan

  6. 6

    Center for Nonlinear and Complex Systems and Department of Physics, Chung-Yuan Christian University, Chungli 32023, Taiwan

  7. 7

    Core Facility for Protein Production and X-Ray Structural Analysis, Academia Sinica, Taipei 11529, Taiwan

  1. †

    Hsin-Liang Chen and Jack C.-C. Hsu contributed equally to this work.

*Institute of Biological Chemistry, Academia Sinica, No. 128, Sec. 2, Academia Rd, Nankang, Taipei 11529, Taiwan

  1. †

    Hsin-Liang Chen and Jack C.-C. Hsu contributed equally to this work.

Publication History

  1. Issue published online: 8 SEP 2010
  2. Article first published online: 23 JUL 2010
  3. Manuscript Accepted: 2 JUL 2010
  4. Manuscript Revised: 24 JUN 2010
  5. Manuscript Received: 7 MAY 2010

Funded by

  • 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

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Keywords:

  • protein folding;
  • RD1;
  • cage;
  • photolabile;
  • photoacoustic calorimetry

Abstract

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.

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