Isothermal titration calorimetry for characterization of recombinant proteins.
Journal
Current opinion in biotechnology
ISSN: 1879-0429
Titre abrégé: Curr Opin Biotechnol
Pays: England
ID NLM: 9100492
Informations de publication
Date de publication:
02 2019
02 2019
Historique:
received:
05
05
2018
revised:
22
06
2018
accepted:
27
06
2018
pubmed:
22
7
2018
medline:
4
7
2019
entrez:
22
7
2018
Statut:
ppublish
Résumé
Isothermal titration calorimetry is widely used to measure the affinities and enthalpies of interaction between proteins and/or small molecules. The quantitative nature of the technique is especially useful in the characterization of recombinant proteins while determining the fraction of protein capable of binding a specific ligand and thus the protein purity. The revealed thermodynamic information sheds light on the binding mechanism, important for the targeted drug design of the biologics. Here we show examples how, together with the thermal shift assay, combination of both techniques enables characterization of protein stability and ligand binding. Furthermore, the binding-linked reactions that strongly affect the observed thermodynamic parameters and must be dissected to obtain the intrinsic parameters that are necessary for the structure-based rational drug design are being demonstrated using inhibitors of Hsp90, an anticancer target protein.
Identifiants
pubmed: 30031160
pii: S0958-1669(18)30040-5
doi: 10.1016/j.copbio.2018.06.003
pii:
doi:
Substances chimiques
Ligands
0
Recombinant Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Review
Langues
eng
Sous-ensembles de citation
IM
Pagination
9-15Informations de copyright
Copyright © 2018. Published by Elsevier Ltd.