| Differential scanning fluorimetry measurement of protein stability changes upon binding to glycosaminoglycans: a screening test for binding specificity. | |
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MedLine Citation:
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PMID: 20353159 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The interaction between glycosaminoglycans (GAGs) and proteins is important for the regulation of protein transport and activity. Here we present a novel method for the measurement of protein-GAG interactions suitable for high-throughput screening, able to discriminate between the interactions of a protein with GAGs of different structures. Binding of proteins to the GAG heparin, a proxy for sulfated regions of extracellular heparan sulfate, was found to enhance the stability of three test proteins, fibroblast growth factors (FGFs)-1, -2, and -18. Chemically modified heparins and heparin oligosaccharides of different lengths stabilized the three FGFs to different extents, depending on the pattern of sugar binding specificity. The method is based on a differential scanning fluorescence approach. It uses a Sypro Orange dye, which binds to exposed core residues of a denatured protein and results in an increased fluorescence signal. It is convenient, requiring low micromolar amounts of protein and ligand compared to other interaction assays, employing only a real-time polymerase chain reaction (PCR) instrument. |
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Authors:
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Katarzyna A Uniewicz; Alessandro Ori; Ruoyan Xu; Yassir Ahmed; Mark C Wilkinson; David G Fernig; Edwin A Yates |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Analytical chemistry Volume: 82 ISSN: 1520-6882 ISO Abbreviation: Anal. Chem. Publication Date: 2010 May |
Date Detail:
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Created Date: 2010-04-30 Completed Date: 2010-08-18 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0370536 Medline TA: Anal Chem Country: United States |
Other Details:
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Languages: eng Pagination: 3796-802 Citation Subset: IM |
Affiliation:
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School of Biological Sciences, University of Liverpool, Liverpool, L69 7ZB, United Kingdom. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Binding Sites Calorimetry, Differential Scanning Fibroblast Growth Factor 1 / chemistry, metabolism Fibroblast Growth Factor 2 / chemistry, metabolism Fluorometry* Glycosaminoglycans / chemistry*, metabolism* Polymerase Chain Reaction Protein Binding Protein Stability |
| Chemical | |
Reg. No./Substance:
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0/Glycosaminoglycans; 103107-01-3/Fibroblast Growth Factor 2; 104781-85-3/Fibroblast Growth Factor 1 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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