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dc.contributor.authorWang, Ting
dc.contributor.authorZhou, Zheng
dc.contributor.authorBunagan, Michelle R.
dc.contributor.authorDu, Degou
dc.contributor.authorBai, Yawen
dc.contributor.authorGai, Feng
dc.identifier.citationWang, T.; Zhou, Z.; Bunagan, M.R.; Du, D.G.; Bai, Y.W.; Gai, F. "Probing the Folding Intermediate of Rd-apocyt b562 by Protein Engineering and Infrared T-Jump," Protein Science, 2007, 16, 1176-1183.en_US
dc.descriptionFile not available for download due to copyright restrictionsen_US
dc.description.abstractSmall proteins often fold in an apparent two-state manner with the absence of detectable early-folding intermediates. Recently, using native-state hydrogen exchange, intermediates that exist after the rate-limiting transition state have been identified for several proteins. However, little is known about the folding kinetics from these post-transition intermediates to their corresponding native states. Herein, we have used protein engineering and a laser-induced temperature-jump (T-jump) technique to investigate this issue and have applied it to Rd-apocyt b562, a four-helix bundle protein. Previously, it has been shown that Rd-apocyt b562 folds via an on-pathway hidden intermediate, which has only the N-terminal helix unfolded. In the present study, a double mutation (V16G/I17A) in the N-terminal helix of Rd-apocyt b562 was made to further increase the relative population of this intermediate state at high temperature by selectively destabilizing the native state. In the circular dichroism thermal melting experiment, this mutant showed apparent two-state folding behavior. However, in the T-jump experiment, two kinetic phases were observed. Therefore, these results are in agreement with the idea that a folding intermediate is populated on the folding pathway of Rd-apocyt b562. Moreover, it was found that the exponential growth rate of the native state from this intermediate state is roughly (25 μsec)−1 at 65°C.en_US
dc.publisherJohn Wiley & Sons, Inc.en_US
dc.titleProbing the Folding Intermediate of Rd-apocyt b562 by Protein Engineering and Infrared T-jumpen_US
prism.publicationNameProtein Scienceen_US

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