The Limited Proteolysis technology coupled to next-generation quantitative mass spectrometry (LiP-MS) is a novel approach that enables the unbiased profiling of structural protein changes across the whole proteome. Structural changes to the proteome can result from a variety of stimuli such as heat shock, protein-protein interactions, compound binding, and posttranslational modifications. The structural changes affect the kinetics of the proteolytic cleavage in the LiP reaction and can be probed using next-generation quantitative proteomics approaches such as HRM.
LiP-MS for Target Deconvolution
LiP-MS has emerged as a label-free technology to identify and analyze changes in protein structures/steric hindrance caused by compound binding in cellular lysates. The underlying concept of LiP-MS is based on proteins being subjected to proteolysis in the presence or absence of a compound. When a compound binds to a protein, it induces a specific structural state in the protein target that can be exploited during proteolysis. This unique structural state changes the pattern of proteolysis and leads to the formation of unique peptides that can be characterized by HRM mass spectrometry.