title: High-throughput bioconjugation for enhanced 193 nm photodissociation via droplet phase initiated ion/ion chemistry using a front-end dual spray reactor.
abstract: Fast online chemical derivatization of peptides with an aromatic label for enhanced 193 nm ultraviolet photodissociation (UVPD) is demonstrated using a dual electrospray reactor implemented on the front-end of a linear ion trap (LIT) mass spectrometer. The reactor facilitates the intersection of protonated peptides with a second population of chromogenic 4-formyl-1,3-benzenedisulfonic acid (FBDSA) anions to promote real-time formation of ion/ion complexes at atmospheric pressure. Subsequent collisional activation of the ion/ion intermediate results in Schiff base formation generated via reaction between a primary amine in the peptide cation and the aldehyde moiety of the FBDSA anion. Utilizing 193 nm UVPD as the subsequent activation step in the MS(3) workflow results in acquisition of greater primary sequence information relative to conventional collision induced dissociation (CID). Furthermore, Schiff-base-modified peptides exhibit on average a 20% increase in UVPD efficiency compared to their unmodified counterparts. Due to the efficiency of covalent labeling achieved with the dual spray reactor, we demonstrate that this strategy can be integrated into a high-throughput LC-MS(n) workflow for rapid derivatization of peptide mixtures.
authors: Department of Chemistry, The University of Texas at Austin , 105 East 24th Street, Austin, Texas 78712, United States.
authors: Victoria C
authors: VC
authors: Cotham
authors: Department of Chemistry, The University of Texas at Austin , 105 East 24th Street, Austin, Texas 78712, United States.
authors: Jared B
authors: JB
authors: Shaw
authors: Department of Chemistry, The University of Texas at Austin , 105 East 24th Street, Austin, Texas 78712, United States.
authors: Jennifer S
authors: JS
authors: Brodbelt
mesh_terms: D000595:Amino Acid Sequence; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry*; D055668:Photochemical Processes; D012545:Schiff Bases / Q000737:chemistry; D013451:Sulfonic Acids / Q000737:chemistry; D014466:Ultraviolet Rays
chemical_list: D010455:Peptides; D012545:Schiff Bases; D013451:Sulfonic Acids