title: Energetics-based methods for protein folding and stability measurements.
abstract: Over the past 15 years, a series of energetics-based techniques have been developed for the thermodynamic analysis of protein folding and stability. These techniques include Stability of Unpurified Proteins from Rates of amide H/D Exchange (SUPREX), pulse proteolysis, Stability of Proteins from Rates of Oxidation (SPROX), slow histidine H/D exchange, lysine amidination, and quantitative cysteine reactivity (QCR). The above techniques, which are the subject of this review, all utilize chemical or enzymatic modification reactions to probe the chemical denaturant- or temperature-induced equilibrium unfolding properties of proteins and protein-ligand complexes. They employ various mass spectrometry-, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)-, and optical spectroscopy-based readouts that are particularly advantageous for high-throughput and in some cases multiplexed analyses. This has created the opportunity to use protein folding and stability measurements in new applications such as in high-throughput screening projects to identify novel protein ligands and in mode-of-action studies to identify protein targets of a particular ligand.
authors.affiliation: Department of Chemistry, Duke University, Durham, North Carolina 27708-0346; email: michael.c.fitzgerald@duke.edu.
authors.forename: M Ariel
authors.initials: MA
authors.lastname: Geer
authors.forename: Michael C
authors.initials: MC
authors.lastname: Fitzgerald
mesh_terms: D000818:Animals; D006801:Humans; D010084:Oxidation-Reduction; D017510:Protein Folding; D055550:Protein Stability; D011506:Proteins / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013816:Thermodynamics
chemical_list: D011506:Proteins
keywords: mass spectrometry; protein H/D exchange; protein folding; proteomics; thermodynamics