Capturing native protein states in real-time using a novel MS-compatible phosphatase and protease inhibitor formulation

摘要

Numerous traditional phosphatase and protease inhibitor cocktails can cause issues during analysis by mass spectrometry necessitating caution with their use. We have derived an MS friendly cocktail which has been designed for maximal protease and phosphatase inhibition containing no irreversible covalent modifiers or substances which would interfere with enrichment or chromatographic methods. Enrichment of phosphopeptides with IMAC was not significantly compromised in the presence of the inhibitor formulation. Testing of the phosphopeptide inhibitor cocktail against a variety of phosphatases was used to demonstrate both the effectiveness of the mixture against a diverse group of enzymes as well as suitability of the components for MS analyses.