Molecular Biology Commons^™

Mass Spectrometry Tools For Analysis Of Intermolecular Interactions, Jared Auclair, Mohan Somasundaran, Karin Green, James Evans, Celia Schiffer, Dagmar Ringe, Gregory Petsko, Jeffrey Agar

Celia A. Schiffer

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.