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Proceedings Paper

Distribution Of Distances In Native And Denatured Troponin I, From Frequency-Domain Measurements Of Fluorescence Energy Transfer
Author(s): Herbert C Cheung; Chien-Kao Wang; Ignacy Gryczynski; Michael L Johnson; Joseph R Lakowicz
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Paper Abstract

We used time-dependent fluorescence energy transfer to determine the distribution of donor-to-acceptor distances in native and denatured troponin I(TnI). The single tryptophan residue (trp 158) of TnI served as the donor (D), and the acceptor (A) was a labeled cysteine residue (cys 133). The time-dependent intensity decays of the donor were measured by the frequency-domain method. The frequency-response of the donor emission, in the absence and presence of acceptor, was used to recover the distribution of D to Å distances, using an algorithm which accounts for the intrinsic multi-exponential decay of the donor. In the native state the D-A distribution is characterized by an average distance of 23 A and a half-width of 12 Å. Denaturation results in a modest increase in the average distance to 27 A, and a dramatic increase in half-width to 47 Å. We believe the ability to recover distance distributions will have numerous applications in the characterization of biological macromolecules.

Paper Details

Date Published: 24 June 1988
PDF: 7 pages
Proc. SPIE 0909, Time-Resolved Laser Spectroscopy in Biochemistry, (24 June 1988); doi: 10.1117/12.945382
Show Author Affiliations
Herbert C Cheung, University of Maryland at Baltimore (United States)
Chien-Kao Wang, University of Maryland at Baltimore (United States)
Ignacy Gryczynski, University of Maryland at Baltimore (United States)
Michael L Johnson, University of Virginia (United States)
Joseph R Lakowicz, University of Alabama at Birmingham (United States)

Published in SPIE Proceedings Vol. 0909:
Time-Resolved Laser Spectroscopy in Biochemistry
Joseph R. Lakowicz, Editor(s)

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