Journal
ANALYTICAL BIOCHEMISTRY
Volume 508, Issue -, Pages 114-117Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2016.05.019
Keywords
Differential scanning fluorimetry; Enzyme kinetics; Enzyme stability; Urea; Native denaturation; Lactate dehydrogenase
Funding
- Natural Sciences and Engineering Research Council (NSERC) of Canada [6793]
- Canada Research Chairs program
- Ontario Graduate Scholarship
Ask authors/readers for more resources
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 (K-nd), and protein unfolding by urea (I-50). 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. (C) 2016 Elsevier Inc. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available