Stable isotope labeling strategy for protein-ligand binding analysis in multi-component protein mixtures.

ABSTRACT: Described here is a stable isotope labeling protocol that can be used with a chemical modification- and mass spectrometry-based protein-ligand binding assay for detecting and quantifying both the direct and indirect binding events that result from protein-ligand binding interactions. The protocol utilizes an H(2) (16)O(2) and H(2) (18)O(2) labeling strategy to evaluate the chemical denaturant dependence of methionine oxidation in proteins both in the presence and absence of a target ligand. The differential denaturant dependence to the oxidation reactions performed in the presence and absence of ligand provides a measure of the protein stability changes that occur as a result of direct interactions of proteins with the target ligand and/or as a result of indirect interactions involving oth

SUBMITTER: DeArmond PD 

PROVIDER: S-EPMC3085011 | biostudies-literature | 2011 Mar

REPOSITORIES: biostudies-literature