End-to-end distance distribution in bradykinin observed by Förster resonance energy transfer.
Förster resonance energy transfer (FRET) was used to study the conformational dynamics of bradykinin related peptides. The fluorescent probe aminobenzoic acid (Abz) bound to the amino terminal of bradykinin maintained its fluorescence characteristics, like high quantum yield and excited state decay dominated by a lifetime of 8.3 ns. The binding of the acceptor group N-[2, 4-dinitrophenyl]-ethylenediamine (EDDnp) to the carboxy terminal of Abz labeled bradykinin resulted in a drastic decrease of the fluorescence intensity and in a fastening of the excited state decay. The change of the decay kinetics to an heterogeneous process, precludes the use of energy transfer models based on a single fixed distance between donor and acceptor. The computational package CONTIN was employed to the analysis of time-resolved fluorescence data, allowing the recovery of a distance distribution between donor and acceptor corresponding to the end-to-end distance of the labeled peptide. The distance distribution reflects the occurrence of distinct conformations for the peptide, that coexist in equilibrium during the fluorescence lifetime. We observed three distance populations for bradykinin in water, that merged to two populations when the solvent was trifluoroethanol (TFE). The results were consistent with those obtained from circular dichroism spectroscopy, that showed structural flexibility in water and the presence of more defined secondary structure in TFE. We also studied several peptides related to bradykinin, and the results emphasized the formation of turns involving the proline residues and the decrease of conformational flexibility induced by using TFE as the solvent.
de Souza
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ES
Instituto de Física da Universidade de São Paulo, Caixa Postal 66318, CEP 05315-970, São Paulo, Brazil.
Hirata
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IY
Juliano
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L
Ito
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AS
D000595:Amino Acid Sequence; D001920:Bradykinin / Q000737:chemistry*; D002942:Circular Dichroism; D004735:Energy Transfer; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes
D005456:Fluorescent Dyes; D001920:Bradykinin