Hydrogen/deuterium exchange- and protease digestion-based screening assay for protein-ligand binding detection.

ABSTRACT: A protease digestion strategy was incorporated into single-point stability of unpurified proteins from rates of H/D exchange (SUPREX), which is a hydrogen/deuterium (H/D) exchange- and mass spectrometry-based assay for the detection of protein-ligand binding. Single-point SUPREX is an abbreviated form of SUPREX in which protein-ligand binding interactions are detected by measuring the increase in a protein's thermodynamic stability upon ligand binding. The new protease digestion protocol provides a noteworthy increase in the efficiency of single-point SUPREX because peptide masses can be determined with greater precision than intact protein masses in the matrix-assisted laser desorption ionization (MALDI) readout of single-point SUPREX. The protocol was evaluated in test screens on two mod

SUBMITTER: Hopper ED 

PROVIDER: S-EPMC2858457 | biostudies-literature | 2009 Aug

REPOSITORIES: biostudies-literature