Abstract
Multiple Reaction Monitoring Initiated Detection and Sequencing (MIDAS™) is a mass spectrometry-based technique for the detection and characterization of specific post-translational modifications (Unwin et al. 4:1134–1144, 2005), for example acetylated lysine residues (Griffiths et al. 18:1423–1428, 2007). The MIDAS™ technique has application for discovery and analysis of acetylation sites. It is a hypothesis-driven approach that requires a priori knowledge of the primary sequence of the target protein and a proteolytic digest of this protein. MIDAS essentially performs a targeted search for the presence of modified, for example acetylated, peptides. The detection is based on the combination of the predicted molecular weight (measured as mass–charge ratio) of the acetylated proteolytic peptide and a diagnostic fragment (product ion of m/z 126.1), which is generated by specific fragmentation of acetylated peptides during collision induced dissociation performed in tandem mass spectrometry (MS) analysis. Sequence information is subsequently obtained which enables acetylation site assignment. The technique of MIDAS was later trademarked by ABSciex for targeted protein analysis where an MRM scan is combined with full MS/MS product ion scan to enable sequence confirmation.
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Acknowledgments
Financial support from Leukaemia Lymphoma Research, UK (ADW, CE, JG), Biotechnology and Biological Sciences Research Council (BC), Cancer Research, UK (ADW, JRG), and NIHR Manchester Biomedical Research Centre (RDU). Financial support from Engineering and Physical Sciences Research Council, ChELSI initiative EP/E036252/1 (CE).
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Evans, C.A., Griffiths, J.R., Unwin, R.D., Whetton, A.D., Corfe, B.M. (2013). Application of the MIDAS Approach for Analysis of Lysine Acetylation Sites. In: Hake, S., Janzen, C. (eds) Protein Acetylation. Methods in Molecular Biology, vol 981. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-305-3_3
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DOI: https://doi.org/10.1007/978-1-62703-305-3_3
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