MT-MAMS: Protein Methyltransferase Motif Analysis by Mass Spectrometry

Journal of Proteome Research
Joshua J HameyMarc R Wilkins

Abstract

Protein methyltransferases often recognize their substrates through linear sequence motifs. The determination of these motifs is critical to understand methyltransferase mechanism, function, and drug targeting. Here we describe MT-MAMS (methyltransferase motif analysis by mass spectrometry), a quantitative approach to characterize methyltransferase substrate recognition motifs. In MT-MAMS, peptide sets are synthesized which contain all amino acid substitutions at single positions within a template sequence. These are then incubated with the methyltransferase of interest in the presence of deuterated S-adenosyl methionine (D3-AdoMet). The use of this heavy methyl donor gives unique mass shifts to methylated peptides, allowing their unambiguous quantification by mass spectrometry. The stoichiometry of methylation resulting from each substitution is then derived, and finally the methyltransferase substrate recognition motif is generated. We validated MT-MAMS by application to lysine methyltransferase G9a, generating the substrate recognition motif (TKRN)-(A > RS > G)-(R ≫ K)-K-(STRCKMAQHG)-Φ; this is highly similar to that previously determined by peptide arrays. We then determined the recognition motif of yeast lysine elongation ...Continue Reading

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Citations

Jul 28, 2020·The Biochemical Journal·Sarah A MannBryan Knuckley
Jul 24, 2020·Molecular Biology and Evolution·Samantha J Emery-CorbinAaron R Jex
May 18, 2021·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Valentina Lukinović, Kyle K Biggar
Oct 24, 2019·Current Protein & Peptide Science·Yogita Jethmalani, Erin M Green
Jan 25, 2020·Current Protein & Peptide Science·Owen M Price, Joan M Hevel
Nov 6, 2021·Frontiers in Molecular Biosciences·Aneta J LenardTobias Madl

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