title: The influence glutamic acid in protonated b3→ b2formation from VGEIG and related analogs.
abstract: A direct pathway for the fragmentation of peptide b3fragment ions to b2ions has, until now, not been identified. Experimental evidence for the formation of a b3anhydride structure and isomerization to an extended macrocycle is demonstrated here by comparison of the completely different fragmentation patterns of the b3ions generated from protonated VGEIG and its methyl ester. In particular, the absence of a b2ion in the fragmentation spectrum of the methyl ester b3indicates that facile fragmentation of an anhydride-type b3is responsible for virtually all b2ions formed. The stability of this b3structure and the ease with which it fragments to the b2may be responsible for the relatively high abundance of the b3and b2ions. IRMPD action spectroscopy measurements indicate the presence of a ring protonated oxazolone in the b2population. VGEIG and three related analogs, VALEIG, VADEIG, and V(Aib)EIG were studied by QCID-HDX-SORI experiments in an FT-ICR instrument, and provide significant evidence for extensive alpha proton scrambling in an ion-molecule complex formed between the b2and neutral loss fragment following formation of the b2. MS3and HDX of VG(2,2-d2)EIG indicate that the scrambled b2ions have the same structure as the unscrambled b2. Based on these data and with the support of molecular modeling, we propose a new mechanism for this scrambling, in which the alpha protons are transferred in a multistep pathway during an ion-molecule complex formed between the b2and amino-terminated anhydride ring neutral loss component.
authors.affiliation: Department of Chemistry and Biochemistry, University of Arizona, Tucson, AZ, United States.
authors.forename: Lindsay
authors.initials: L
authors.lastname: Morrison
authors.forename: Arpád
authors.initials: A
authors.lastname: Somogyi
authors.forename: Vicki H
authors.initials: VH
authors.lastname: Wysocki
keywords: Alpha proton scrambling; Anhydride; Peptide fragmentation; b3 ion