Native denaturation differential scanning fluorimetry: determining the effect of urea using a quantitative real-time thermocycler

Childers, C. L., Green, S. R., Dawson, N. J. and Storey, K. B. (2016) Native denaturation differential scanning fluorimetry: determining the effect of urea using a quantitative real-time thermocycler. Analytical Biochemistry, 508, pp. 114-117. (doi: 10.1016/j.ab.2016.05.019) (PMID:27296634)

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Abstract

The effect of protein stability on kinetic function is monitored with many techniques that often require large amounts of expensive substrates and specialized equipment not universally available. We present differential scanning fluorimetry (DSF), a simple high-throughput assay performed in real-time thermocyclers, as a technique for analysis of protein unfolding. Furthermore, we demonstrate a correlation between the half-maximal rate of protein unfolding (Knd), and protein unfolding by urea (I50). This demonstrates that DSF methods can determine the structural stability of an enzyme's active site and can compare the relative structural stability of homologous enzymes with a high degree of sequence similarity.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Dawson, Dr Neal
Authors: Childers, C. L., Green, S. R., Dawson, N. J., and Storey, K. B.
College/School:College of Medical Veterinary and Life Sciences > Institute of Biodiversity Animal Health and Comparative Medicine
College of Medical Veterinary and Life Sciences > School of Biodiversity, One Health & Veterinary Medicine
Journal Name:Analytical Biochemistry
Publisher:Elsevier
ISSN:0003-2697
ISSN (Online):1096-0309
Published Online:11 June 2016

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