title: Amine-reactive neutron-encoded labels for highly plexed proteomic quantitation.
abstract: We describe a novel amine-reactive chemical label that exploits differential neutron-binding energy between (13)C and (15)N isotopes. These neutron-encoded (NeuCode) chemical labels enable up to 12-plex MS1-based protein quantification. Each structurally identical, but isotopically unique, tag is encoded with a 12.6-mDa mass difference-relative to its nearest neighbor-so that peptides bearing these NeuCode signatures do not increase spectral complexity and are detected only upon analysis with very high mass-resolving powers. We demonstrate that the method provides quantitative performance that is comparable to both metabolic labeling and isobaric tagging while combining the benefits of both strategies. Finally, we employ the tags to characterize the proteome of Saccharomyces cerevisiae during the diauxic shift, a metabolic transition from fermentation to aerobic respiration.
authors.affiliation: Department of Biomolecular Chemistry, University of Wisconsin, 420 Henry Mall, Madison, Wisconsin 53706;
authors.forename: Alexander S
authors.initials: AS
authors.lastname: Hebert
authors.forename: Anna E
authors.initials: AE
authors.lastname: Merrill
authors.forename: Jonathan A
authors.initials: JA
authors.lastname: Stefely
authors.forename: Derek J
authors.initials: DJ
authors.lastname: Bailey
authors.forename: Craig D
authors.initials: CD
authors.lastname: Wenger
authors.forename: Michael S
authors.initials: MS
authors.lastname: Westphall
authors.forename: David J
authors.initials: DJ
authors.lastname: Pagliarini
authors.forename: Joshua J
authors.initials: JJ
authors.lastname: Coon
mesh_terms: D000332:Aerobiosis; D000588:Amines / Q000737:chemistry; D002247:Carbon Isotopes; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005285:Fermentation; D007553:Isotope Labeling / Q000379:methods*; D009502:Neutrons; D009587:Nitrogen Isotopes; D040901:Proteomics / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000254:growth & development / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D000588:Amines; D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D029701:Saccharomyces cerevisiae Proteins