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.
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Acknowledgements
The European Commission (Marie Curie Early Stage Training Fellowships) is thanked for funding (KA and AO) and North West Cancer Research Fund for support (DGF).
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Uniewicz, K.A. et al. (2012). Following Protein–Glycosaminoglycan Polysaccharide Interactions with Differential Scanning Fluorimetry. In: Rédini, F. (eds) Proteoglycans. Methods in Molecular Biology, vol 836. Humana Press. https://doi.org/10.1007/978-1-61779-498-8_12
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DOI: https://doi.org/10.1007/978-1-61779-498-8_12
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