Studying submicrosecond protein folding kinetics using a photolabile caging strategy and time-resolved photoacoustic calorimetry
Resource
PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS, 78(14), 2973-2983
Journal
Proteins Structure Function and Bioinformatics
Pages
2973-2983
Date Issued
2010
Date
2010
Author(s)
Chen, Hsin-Liang
Hsu, Jack C.-C.
Viet, Man Hoang
Li, Mai Suan
Hu, Chin-Kun
Liu, Chia-Hsun
Luh, Frederick Y.
Chen, Silvia S.-W.
Chang, Evan S.-H.
Wang, Andrew H.-J.
Hsu, Min-Feng
Fann, Wunshain
Chen, Rita P.-Y.
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.
Type
journal article
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