Photo-induced riboflavin binding to the tryptophan residues of bovine and human serum albumins.
The photobinding between riboflavin and the Trp residues from human and bovine serum albumins at two pH-dependent protein conformations was studied. At pH 7.0 both proteins showed photo-adduct formation with hyperbolic kinetics. In the bovine serum albumin this is attributed to the different locations of the two Trp residues. In the case of the human serum albumin, which has only one Trp residue, this behaviour may be related to different molecular conformations of the protein, as is also manifest in the iodide quenching experiments. At pH 3.5, the kinetics of the photo-adduct formation were found to be slower and showed a monophasic behaviour. These results are due to the conformational change of these proteins at acidic pH; the Trp residues of both proteins being now located in a more hydrophobic environment. When bovine serum albumin was anaerobically irradiated at pH 7.0 in the presence of 14C-riboflavin and then cleaved by CNBr, two peptides were obtained, containing the Trp-134 and Trp-212 residues, respectively. The incorporation of 14C-riboflavin in these samples was significantly higher at the level of the peptide containing the Trp-134 residue. Furthermore, it was demonstrated, that the energy transfer from enzymatically generated triplet acetone to riboflavin can also promote the binding of this vitamin to the Trp residues of human and bovine serum albumins.
Tapia
G
G
Pontificia Universidad Católica de Chile, Facultad de Química, Santiago.
Silva
E
E
D000818:Animals; D001665:Binding Sites; D002417:Cattle; D006801:Humans; D007700:Kinetics; D008027:Light; D011487:Protein Conformation; D012256:Riboflavin / Q000378:metabolism* / Q000494:pharmacology; D012709:Serum Albumin / Q000378:metabolism / Q000528:radiation effects*; D012710:Serum Albumin, Bovine / Q000378:metabolism / Q000528:radiation effects*; D013050:Spectrometry, Fluorescence; D013997:Time Factors; D014364:Tryptophan
D012709:Serum Albumin; D012710:Serum Albumin, Bovine; D014364:Tryptophan; D012256:Riboflavin