Accurate protein-peptide titration experiments by nuclear magnetic resonance using low-volume samples.
Fiche publication
Date publication
janvier 2015
Auteurs
Membres identifiés du Cancéropôle Est :
Dr KIEFFER Bruno
Tous les auteurs :
Kohler C, Recht R, Quinternet M, de Lamotte F, Delsuc MA, Kieffer B
Lien Pubmed
Résumé
NMR spectroscopy allows measurements of very accurate values of equilibrium dissociation constants using chemical shift perturbation methods, provided that the concentrations of the binding partners are known with high precision and accuracy. The accuracy and precision of these experiments are improved if performed using individual capillary tubes, a method enabling full automation of the measurement. We provide here a protocol to set up and perform these experiments as well as a robust method to measure peptide concentrations using tryptophan as an internal standard.
Référence
Methods Mol Biol. 2015;1286:279-96