Distribution Of Distances In Native And Denatured Troponin I, From Frequency-Domain Measurements Of Fluorescence Energy Transfer

Herbert C Cheung; Chien-Kao Wang; Ignacy Gryczynski; Michael L Johnson; Joseph R Lakowicz

doi:10.1117/12.945382

24 June 1988 Distribution Of Distances In Native And Denatured Troponin I, From Frequency-Domain Measurements Of Fluorescence Energy Transfer

Herbert C Cheung, Chien-Kao Wang, Ignacy Gryczynski, Michael L Johnson, Joseph R Lakowicz

Author Affiliations +

Proceedings Volume 0909, Time-Resolved Laser Spectroscopy in Biochemistry; (1988) https://doi.org/10.1117/12.945382
Event: 1988 Los Angeles Symposium: O-E/LASE '88, 1988, Los Angeles, CA, United States

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.

Citation Download Citation

Herbert C Cheung, Chien-Kao Wang, Ignacy Gryczynski, Michael L Johnson, and Joseph R Lakowicz "Distribution Of Distances In Native And Denatured Troponin I, From Frequency-Domain Measurements Of Fluorescence Energy Transfer", Proc. SPIE 0909, Time-Resolved Laser Spectroscopy in Biochemistry, (24 June 1988); https://doi.org/10.1117/12.945382

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