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Conformational Dynamics of a Fast Folding Cytochrome Captured by Time-Resolved Spectroscopy

Bouley Ford, Nicole D. and Shin, Dong Woo and Gray, Harry B. and Winkler, Jay R. (2014) Conformational Dynamics of a Fast Folding Cytochrome Captured by Time-Resolved Spectroscopy. Biophysical Journal, 106 (2). 473A. ISSN 0006-3495. https://resolver.caltech.edu/CaltechAUTHORS:20140710-082331092

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Abstract

We probe intrachain contact dynamics in unfolded cytochrome cb562 by monitoring heme quenching of excited ruthenium photosensitizers covalently bound to residues along the polypeptide. Tertiary contact formation kinetics provide insight into the upper "speed limit" for protein folding rates. The rate constants exhibit a power-law dependence on the number of peptide bonds between the heme and Ru complex. Adherence of our data to a slope of −1.5 is consistent with theoretical models for ideal, freely-jointed Gaussian chain polymers, but its magnitude is smaller than reported for synthetic polypeptides. We also examine rates of contact formation within a stable loop in a His63-heme ligated form of the protein. Additionally, we resolve millisecond-timescale folding by coupling time-resolved fluorescence energy transfer (trFRET) to a continuous flow microfluidic mixer to obtain intramolecular distance distributions throughout the folding process. Our results suggest that cytochrome cb562 is minimally frustrated.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1016/j.bpj.2013.11.2674DOIArticle
http://www.sciencedirect.com/science/article/pii/S0006349513039313PublisherArticle
http://www.biophysics.org/2014meeting/Main/tabid/4177/Default.aspxOrganizationConference Website
ORCID:
AuthorORCID
Gray, Harry B.0000-0002-7937-7876
Winkler, Jay R.0000-0002-4453-9716
Additional Information:© 2014 Biophysical Society. Published by Elsevier Inc.
Issue or Number:2
Record Number:CaltechAUTHORS:20140710-082331092
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20140710-082331092
Official Citation:Nicole Bouley Ford, Dong Woo Shin, Harry B. Gray, Jay R. Winkler, Conformational Dynamics of a Fast Folding Cytochrome Captured by Time-Resolved Spectroscopy, Biophysical Journal, Volume 106, Issue 2, Supplement 1, 28 January 2014, Page 473a, ISSN 0006-3495, http://dx.doi.org/10.1016/j.bpj.2013.11.2674. (http://www.sciencedirect.com/science/article/pii/S0006349513039313)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:47117
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:10 Jul 2014 16:10
Last Modified:22 Nov 2019 09:58

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