Mass spectrometry tools for analysis of intermolecular interactions
Department of Pediatrics; Department of Biochemistry and Molecular Pharmacology
Mass Spectrometry; Protein Binding; Binding Sites
Biochemistry, Biophysics, and Structural Biology | Molecular Biology | Pharmacology, Toxicology and Environmental Health
The small quantities of protein required for mass spectrometry (MS) make it a powerful tool to detect binding (protein-protein, protein-small molecule, etc.) of proteins that are difficult to express in large quantities, as is the case for many intrinsically disordered proteins. Chemical cross-linking, proteolysis, and MS analysis, combined, are a powerful tool for the identification of binding domains. Here, we present a traditional approach to determine protein-protein interaction binding sites using heavy water ((18)O) as a label. This technique is relatively inexpensive and can be performed on any mass spectrometer without specialized software.
DOI of Published Version
Methods Mol Biol. 2012;896:387-98. Link to article on publisher's site
Methods in molecular biology (Clifton, N.J.)
Auclair, Jared R.; Somasundaran, Mohan; Green, Karin M.; Evans, James E.; Schiffer, Celia A.; Ringe, Dagmar; Petsko, Gregory A.; and Agar, Jeffrey N., "Mass spectrometry tools for analysis of intermolecular interactions" (2012). Biochemistry and Molecular Pharmacology Publications and Presentations. 153.