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Conformational dynamics of a fast folding cytochrome captured by electron transfer, time-resolved fluorescence energy transfer, and microfluidic mixing

Ford, Nicole B. and Shin, Dong Woo and Gray, Harry B. and Winkler, Jay R. (2013) Conformational dynamics of a fast folding cytochrome captured by electron transfer, time-resolved fluorescence energy transfer, and microfluidic mixing. In: 245th ACS National Meeting & Exposition, Abstracts of Papers, April 7-11, 2013, New Orleans, LA. https://resolver.caltech.edu/CaltechAUTHORS:20131010-083823832

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Abstract

Cytochrome c-b_(562) belongs to an interesting family of four-helix bundle cytochromes that have nearly identical structures yet widely divergent folding pathways, creating a unique opportunity to study the relationship between primary sequences and folding in proteins with similar topologies. We characterize the dynamics of cytochrome c-b_(562) through a combination of electron transfer and fluorescence energy transfer (FRET). The dynamics of intrachain diffusion in unfolded cytochromes are probed by monitoring contact quenching of a photoexcited, covalently-bound [Ru(bpy)2(IA-phen)]^(2+) dye by the heme Fe^(III). Quenching of the *Ru^(II) luminescence decay provides information about the electron transfer rates, which correspond to the rates of transient contact formation. We resolve fast folding by coupling time-resolved FRET with an ultrafast continuous flow mixer. FRET occurs between a small, covalently-bound dansyl fluorophore and the heme. Fitting the dansyl fluorescence decay following picosecond laser excitation provides us with time-dependent dansyl-heme distance distributions during the two-state folding process.


Item Type:Conference or Workshop Item (Paper)
Related URLs:
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http://presentations.acs.org/common/tracks.aspx/Spring2013OrganizationUNSPECIFIED
ORCID:
AuthorORCID
Winkler, Jay R.0000-0002-4453-9716
Additional Information:© 2013 American Chemical Society.
Record Number:CaltechAUTHORS:20131010-083823832
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20131010-083823832
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:41851
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:10 Oct 2013 18:50
Last Modified:03 Oct 2019 05:52

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