Dynamics of the Lens culinaris agglutinin-lactotransferrin and serotransferrin complexes, followed by fluorescence intensity quenching of fluorescein (FITC) with iodide and temperature

Dynamics of the fluorescent Lens culinaris agglutinin-fluorescein complex (LCA-FITC) are studied in absence and in presence of two glycoproteins, lactotransferrin (LTF) and serotransferrin (STF). Glycans of the serotransferrin are not fucosylated, while those of the lactotransferrin have an alpha-1,6-fucose bound to the N-acetylglucosamine residue linked to the peptide chain, and an alpha-1,3-fucose bound to the N-acetyllactosamine residues. Interaction between the lectin and the two glycoproteins occurs via the carbohydrate residues. Affinity between LCA and LTF is 50 times higher than that between LCA and STF, as a result of the alpha-1,6-fucose-LCA linkage. In the present work, we studied the effect of the tight bond between the alpha-1,6-fucose and LCA on the dynamics of the amino acids of the lectin, by fluorescence intensity quenching with iodide and by thermal intensity quenching. Fluorescence intensity quenching with iodide indicates that the bimolecular diffusion constant of iodide is 2.402 +/- 0.068 x 10(9) and 1.160 +/- 0.090 x 10(9) M-1 s(-1), when the interaction occurs with free fluorescein and with fluorescein bound to LCA, respectively. Binding of STF or LTF to the LCA-FITC complex yields a bimolecular diffusion constant of 1.675 +/- 0.06 x 10(9) and 1.155 +/- 0.087 x 10(9) M-1 s(-1), respectively. Thermal intensity quenching does not occur for fluorescein free in solution while it is linear with temperature with a relative change of 0.656%, 0.889% and 0.488% for FITC-LCA, FITC-LCA-LTF and FITC-LCA-STF complexes, respectively. Fluorescence intensity quenching with iodide and thermal quenching experiments indicate that the dynamics of LCA increase as the result of the flexibility of the carbohydrate residues (case of STF-LCA complex), and the presence of the a-1,6-fucose inhibits the effect of the other carbohydrate residues as the result of the tight bond that exists between the fucose and the lectin (case of LTF-LCA complex). (C) 1998 Elsevier Science B.V. All rights reserved.