Premium
Looking for protein stabilizing drugs with thermal shift assay
Author(s) -
Andreotti Giuseppina,
Monticelli Maria,
Cubellis Maria Vittoria
Publication year - 2015
Publication title -
drug testing and analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.065
H-Index - 54
eISSN - 1942-7611
pISSN - 1942-7603
DOI - 10.1002/dta.1798
Subject(s) - small molecule , chemistry , mutant , high throughput screening , fabry disease , enzyme , chromatography , pharmacology , computational biology , biochemistry , biology , medicine , disease , gene
Thermal shift assay can be used for the high‐throughput screening of pharmacological chaperones. These drugs are small molecules that bind a mutant protein and stabilize it. We demonstrated the robustness, reproducibility and versatility of the method using two molecules that are in clinical trial for Fabry or Pompe disease, Deoxygalactonojirimycin and N‐Butyldeoxynojirimycin, and their target enzymes, lysosomal alpha‐galactosidaseA and alpha‐glucosidase, as test cases. We assessed the influence of solvents and of scanning rate on the measures. We showed that a value that is equivalent to the melting temperature can be obtained by the first derivatives of raw data. We discuss the advantages of the method and the precaution to be taken in running the experiments. © 2015 The Authors Drug Testing and Analysis Published by John Wiley & Sons Ltd.