The effect of resonance energy homotransfer on the intrinsic tryptophan fluorescence emission of the bothropstoxin-I dimer.
Bothopstoxin-I (BthTX-I) is a homodimeric Lys49-PLA2 homologue from the venom of Bothrops jararacussu in which a single Trp77 residue is located at the dimer interface. Intrinsic tryptophan fluorescence emission (ITFE) quenching by iodide and acrylamide has confirmed that a dimer to monomer transition occurs on reducing the pH from 7.0 to 5.0. Both the monomer and the dimer showed an excitation wavelength-dependent increase in the fluorescence emission maximum, however the excitation curve of the dimer was blue-shifted with respect to the monomeric form. No differences in the absorption or circular dichroism spectra between pH 5.0 and 7.0 were observed, suggesting that this curve shift is due neither to altered electronic ground states nor to exciton coupling of the Trp residues. We suggest that fluorescence resonance energy homotransfer between Trp77 residues at the BthTX-I dimer interface results in excitation of an acceptor Trp population which demonstrates a red-shifted fluorescence emission.
de Oliveira
A H
AH
Department of Biochemistry and Immunology, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto-SP, Brazil.
Giglio
J R
JR
Andrião-Escarso
S H
SH
Ward
R J
RJ
D000818:Animals; D017837:Bothrops; D003435:Crotalid Venoms / Q000737:chemistry*; D019281:Dimerization; D004735:Energy Transfer; D006863:Hydrogen-Ion Concentration; D007454:Iodides; D008958:Models, Molecular; D017433:Protein Structure, Secondary; D013050:Spectrometry, Fluorescence / Q000379:methods; D014364:Tryptophan
D003435:Crotalid Venoms; D007454:Iodides; C078097:bothropstoxin; D014364:Tryptophan