title: Glycosaminoglycan-protein interaction studies using fluorescence spectroscopy.
abstract: Fluorescence spectroscopy is a quantitative analytical tool that has been extensively used to provide structural and dynamical information on GAG-protein complexes. It possesses major advantages including high sensitivity, relative ease of applicability, and wide range of available fluorescence labels and probes. It has been applied to practically every protein-GAG system through the use of either intrinsic (e.g., Trp) or extrinsic (e.g., a non-covalent fluorophore) probe. For studies involving GAGs, it forms the basis for measurement of dissociation constant of complexes and the stoichiometry of binding, which helps elucidate many other thermodynamic and/or mechanistic parameters. We describe the step-by-step procedure to measure the affinity of GAG-protein complexes, parse the ionic and nonionic components of the free energy of binding, and identify the site of GAG binding through competitive binding experiments.
authors.affiliation: Department of Medicinal Chemistry and Institute for Structural Biology and Drug Discovery, Virginia Commonwealth University, 800 E. Leigh Street, Suite 212, Richmond, VA, 23219, USA.
authors.forename: Rio S
authors.initials: RS
authors.lastname: Boothello
authors.forename: Rami A
authors.initials: RA
authors.lastname: Al-Horani
authors.forename: Umesh R
authors.initials: UR
authors.lastname: Desai
mesh_terms: D000991:Antithrombins / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry; D006025:Glycosaminoglycans / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism; D007477:Ions; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D000991:Antithrombins; D005456:Fluorescent Dyes; D006025:Glycosaminoglycans; D007477:Ions; D011506:Proteins; D006493:Heparin