title: Quantitative Analysis and Discovery of Lysine and Arginine Modifications.
abstract: Post-translational modifications (PTMs) affect protein function, localization, and stability, yet very little is known about the ratios of these modifications. Here, we describe a novel method to quantitate and assess the relative stoichiometry of Lys and Arg modifications (QuARKMod) in complex biological settings. We demonstrate the versatility of this platform in monitoring recombinant protein modification of peptide substrates, PTMs of individual histones, and the relative abundance of these PTMs as a function of subcellular location. Lastly, we describe a product ion scanning technique that offers the potential to discover unexpected and possibly novel Lys and Arg modifications. In summary, this approach yields accurate quantitation and discovery of protein PTMs in complex biological systems without the requirement of high mass accuracy instrumentation.
authors.forename: James J
authors.identifier: 0000-0002-5612-0680
authors.initials: JJ
authors.lastname: Galligan
authors.forename: Philip J
authors.initials: PJ
authors.lastname: Kingsley
authors.forename: Orrette R
authors.initials: OR
authors.lastname: Wauchope
authors.forename: Michelle M
authors.initials: MM
authors.lastname: Mitchener
authors.forename: Jeannie M
authors.identifier: 0000-0001-9699-0931
authors.initials: JM
authors.lastname: Camarillo
authors.forename: James A
authors.initials: JA
authors.lastname: Wepy
authors.affiliation: Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado , Anschutz Medical Campus, Aurora, Colorado 80045, United States.
authors.forename: Peter S
authors.identifier: 0000-0003-0324-0862
authors.initials: PS
authors.lastname: Harris
authors.affiliation: Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado , Anschutz Medical Campus, Aurora, Colorado 80045, United States.
authors.forename: Kristofer S
authors.identifier: 0000-0002-1499-6174
authors.initials: KS
authors.lastname: Fritz
authors.forename: Lawrence J
authors.identifier: 0000-0002-7834-6285
authors.initials: LJ
authors.lastname: Marnett
mesh_terms: D001120:Arginine / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D057809:HEK293 Cells; D006657:Histones / Q000737:chemistry*; D006801:Humans; D006868:Hydrolysis; D056484:Jumonji Domain-Containing Histone Demethylases / Q000737:chemistry; D008239:Lysine / Q000032:analysis*; D010455:Peptides / Q000737:chemistry*; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D006657:Histones; D010455:Peptides; D011994:Recombinant Proteins; D001120:Arginine; D056484:Jumonji Domain-Containing Histone Demethylases; D008239:Lysine