Fluorescence Energy-Transfer Probes of Conformation in Peptides: The 2-Aminobenzamide/Nitrotyrosine Pair

Jens Øllgaard Duus, Morten Meldal, Jay R. Winkler

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Abstract

The photophysical properties of the fluorescence energy-transfer donor, 2-aminobenzamide (Abz), and nitrotyrosine (Tyr-NO2) acceptor have been evaluated. The 31-Å Fo¨rster radius for this donor-acceptor pair makes it useful for evaluating molecular separations in the range of 15-45 Å. Abz and Tyr-NO2 have been incorporated into four medium-sized synthetic peptides (1, Abz-SEEEEKKKKEEEEKKKK(Tyr(NO2))D; 2, Abz-AEAAAKHAAAHEAAAKA(Tyr(NO2))D; 3, Abz-FAQKEPAFLKEYHLL(Tyr(NO2))D; 4, Abz- LKELKDKLKELKDKLK(Tyr(NO2))LKD). Nonexponential Abz fluorescence decays indicate that the synthetic peptides do not assume single, static conformations in solution. The fluorescence decay kinetics in three of the four peptides have been fit to a model in which the peptide conformations are described by Gaussian distributions of donor-acceptor distances centered at rC (rC (fwhm): 2, 27.0 (8.1); 3, 25.6 (12.9); 4, 21.1 (22.1) Å). Values of rC are in good agreement with distances obtained from molecular dynamics simulations (rMD: 2, 28; 3, 21; 4, 22 Å).
Original languageEnglish
JournalJournal of physical chemistry b
Volume102
Pages (from-to)6413-6418
ISSN1520-6106
Publication statusPublished - 1998
Externally publishedYes

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