title: Dynamics of carbohydrate residues of alpha 1-acid glycoprotein (orosomucoid) followed by red-edge excitation spectra and emission anisotropy studies of Calcofluor White.
abstract: Dynamics studies on Calcofluor White bound to the carbohydrate residues of sialylated and asialylated alpha 1-acid glycoprotein (orosomucoid) have been performed. The interaction between the fluorophore and the protein was found to occur preferentially with the glycan residues with a dependence on their spatial conformation. In the presence of sialylated alpha 1-acid glycoprotein, excitation at the red edge of the absorption spectrum of calcofluor does not lead to a shift in the fluorescence emission maximum (440 nm) of the fluorophore. Thus, the emission of calcofluor occurs from a relaxed state. This is confirmed by anisotropy studies as a function of temperature (Perrin plot). In the presence of asialylated alpha 1-acid glycoprotein, red-edge excitation spectra show an important shift (8 nm) of the fluorescence emission maximum of the probe. This reveals that emission of calcofluor occurs before relaxation of the surrounding carbohydrate residues occurs. Emission from a non-relaxed state means that Calcofluor molecules are bound tightly to the carbohydrate residues, a result confirmed by anisotropy studies.
authors.affiliation: Laboratoire de Biophysique Moléculaire, Université des Sciences et Technologies de Lille, Villeneuve d'Ascq, France. jihad-rene.albani@univ-lille1.fr
authors.forename: J R
authors.initials: JR
authors.lastname: Albani
authors.forename: A
authors.initials: A
authors.lastname: Sillen
authors.forename: B
authors.initials: B
authors.lastname: Coddeville
authors.forename: Y D
authors.initials: YD
authors.lastname: Plancke
authors.forename: Y
authors.initials: Y
authors.lastname: Engelborghs
mesh_terms: D001557:Benzenesulfonates / Q000737:chemistry*; D002236:Carbohydrate Conformation; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes / Q000737:chemistry; D015394:Molecular Structure; D009961:Orosomucoid / Q000737:chemistry*; D011485:Protein Binding; D012794:Sialic Acids / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013696:Temperature
chemical_list: D001557:Benzenesulfonates; D005456:Fluorescent Dyes; D009961:Orosomucoid; D012794:Sialic Acids; C007061:C.I. Fluorescent Brightening Agent 28