title: Incorporation of a Reporter Peptide in FPOP Compensates for Adventitious Scavengers and Permits Time-Dependent Measurements.
abstract: Incorporation of a reporter peptide in solutions submitted to fast photochemical oxidation of proteins (FPOP) allows for the correction of adventitious scavengers and enables the normalization and comparison of time-dependent results. Reporters will also be useful in differential experiments to control for the inclusion of a radical-reactive species. This incorporation provides a simple and quick check of radical dosage and allows comparison of FPOP results from day-to-day and lab-to-lab. Use of a reporter peptide in the FPOP workflow requires no additional measurements or spectrometers while building a more quantitative FPOP platform. It requires only measurement of the extent of reporter-peptide modification in a LC/MS/MS run, which is performed by using either data-dependent scanning or an inclusion list. Graphical Abstract ᅟ.
authors.affiliation: Department of Chemistry, Washington University in St. Louis, St. Louis, MO, 63130, USA.
authors.forename: Ben
authors.initials: B
authors.lastname: Niu
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA, 01605, USA.
authors.forename: Brian C
authors.initials: BC
authors.lastname: Mackness
authors.affiliation: Department of Chemistry, Washington University in St. Louis, St. Louis, MO, 63130, USA.
authors.forename: Don L
authors.initials: DL
authors.lastname: Rempel
authors.affiliation: Department of Chemistry, Washington University in St. Louis, St. Louis, MO, 63130, USA.
authors.forename: Hao
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Chemistry, Washington University in St. Louis, St. Louis, MO, 63130, USA.
authors.forename: Weidong
authors.initials: W
authors.lastname: Cui
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA, 01605, USA.
authors.forename: C Robert
authors.initials: CR
authors.lastname: Matthews
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA, 01605, USA.
authors.forename: Jill A
authors.initials: JA
authors.lastname: Zitzewitz
authors.affiliation: Department of Chemistry, Washington University in St. Louis, St. Louis, MO, 63130, USA. mgross@wustl.edu.
authors.forename: Michael L
authors.initials: ML
authors.lastname: Gross
mesh_terms: D002853:Chromatography, Liquid; D016166:Free Radical Scavengers / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D009154:Mutation; D010455:Peptides / Q000737:chemistry*; D055668:Photochemical Processes; D011487:Protein Conformation; D000072105:Superoxide Dismutase-1 / Q000737:chemistry / Q000235:genetics; D057188:Workflow
chemical_list: D016166:Free Radical Scavengers; D010455:Peptides; D000072105:Superoxide Dismutase-1
keywords: Adventitious radical scavenger; FPOP; Mass spectrometry; Reporter; SOD1