title: Following protein-glycosaminoglycan polysaccharide interactions with differential scanning fluorimetry.
abstract: Studies of the structural changes invoked in proteins by the binding of the glycosaminoglycan (GAG) polysaccharide portion of proteoglycans are of increasing importance to research in a wide range of fields, from biochemistry and molecular biology to biotechnology and medicine. One important aspect is the degree of stabilisation or destabilisation induced in a protein by the binding of these anionic materials, and this can affect enzyme activity, the stability of complexes, folding and the formation of aggregates, including those in neurodegenerative processes. A simple method, able to determine the effect of interactions with GAG polysaccharides on protein stability is described, based on the propensity of a fluorescent dye-Sypro™ Orange-to present differentiable fluorescence emission spectra following contact with exposed core amino acid residues. The method requires only commonly available and inexpensive equipment and is suitable for a multi-well format, allowing multiple readings to be made simultaneously.
authors.affiliation: Institute of Integrative Biology, University of Liverpool, Liverpool, UK.
authors.forename: Katarzyna A
authors.initials: KA
authors.lastname: Uniewicz
authors.forename: Alessandro
authors.initials: A
authors.lastname: Ori
authors.forename: Timothy R
authors.initials: TR
authors.lastname: Rudd
authors.forename: Marco
authors.initials: M
authors.lastname: Guerrini
authors.forename: Mark C
authors.initials: MC
authors.lastname: Wilkinson
authors.forename: David G
authors.initials: DG
authors.lastname: Fernig
authors.forename: Edwin A
authors.initials: EA
authors.lastname: Yates
mesh_terms: D005470:Fluorometry / Q000379:methods*; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry*; D011506:Proteins / Q000032:analysis* / Q000737:chemistry*
chemical_list: D011134:Polysaccharides; D011506:Proteins