Energetics-based methods for protein folding and stability measurements.

Published

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