Screening Method for Isopeptides from Small Ubiquitin-Related Modifier-Conjugated Proteins by Ion Mobility Mass Spectrometry

摘要

1. The combination of dual chymotryspin/trypsin proteolysis, solution conditions that promote higher charge states, and IMMS provides a simple, robust and efficient screening method for possible isopeptides from SUMO conjugates. 2. These potential isopeptides can be extracted from the drift time total ion chromatogram as ions with z ≥ +3 and confirmed by tandem mass spectrometry. 3. This method is suitable for all isoforms of human SUMO: SUMO-1. -2, -3, and -4 with only minor adjustment. 4. The technique presented here is meant to complement and facilitate already existing methods for SUMOylation analysis.