Energetics-based methods for protein folding and stability measurements.
Journal Article (Review)
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.
Full Text
Duke Authors
Cited Authors
- Geer, MA; Fitzgerald, MC
Published Date
- January 2014
Published In
Volume / Issue
- 7 /
Start / End Page
- 209 - 228
PubMed ID
- 24896313
Pubmed Central ID
- 24896313
Electronic International Standard Serial Number (EISSN)
- 1936-1335
International Standard Serial Number (ISSN)
- 1936-1327
Digital Object Identifier (DOI)
- 10.1146/annurev-anchem-071213-020024
Language
- eng