Effect of Peptide Fragment Size on the Propensity of Cyclization in Collision-Induced Dissociation: Oligoglycine b(2)-b(8)

TitleEffect of Peptide Fragment Size on the Propensity of Cyclization in Collision-Induced Dissociation: Oligoglycine b(2)-b(8)
Publication TypeJournal Article
Year of Publication2009
AuthorsX. Chen, L. Yu, J.D Steill, J. Oomens, N.C Polfer
JournalJournal of the American Chemical Society
Volume131
Number51
Pagination18272-18282
Date PublishedDec
Type of ArticleArticle
ISBN Number0002-7863
Accession NumberISI:000273615800038
KeywordsAB-INITIO, AMINO-ACIDS, CONFORMATIONS, DEUTERIUM-EXCHANGE-REACTIONS, GAS-PHASE, H/D EXCHANGE, HYDROGEN/DEUTERIUM EXCHANGE, INFRARED-SPECTROSCOPY, MASS-SPECTROMETRY, MULTIPLE-PHOTON DISSOCIATION, PROTONATED TRYPTIC PEPTIDES, TANDEM
Abstract

The chemistry of peptide fragmentation by collision-induced dissociation (CID) is currently being reviewed, as a result of observations that the amino acid sequence of peptide fragments can change upon activation. This rearrangement mechanism is thought to be due to a head-to-tail cyclization reaction, where the N-terminal and C-terminal part of the fragment are fused into a macrocycle (=cyclic peptide) structure, thus "losing" the memory of the original sequence. We present a comprehensive study for a series of b fragment ions, from b(2) to b(8), based on the simplest amino acid residue glycine, to investigate the effect of peptide chain length on the appearance of macrocycle fragment structures. The CID product ions are structurally characterized with a range of gas-phase techniques, including isotope labeling, infrared photodissociation spectroscopy, gas-phase hydrogen/deuterium exchange (using CH3OD), and computational structure approaches. The combined insights from these results yield compelling evidence that smaller b(n) fragments (n = 2, 3) exclusively adopt oxazolone-type structures, whereas a mixture of oxazolone and macrocycle b fragment structures are formed for midsized b(n) fragments, where n = 4-7. As each of these chemical structures exchanges at different rates, it is possible to approximate the relative abundances using kinetic fits to the H/D exchange data. Under the conditions used here, the "slow"-exchanging macrocycle structure represents similar to 30% of the b ion population for b(6)-b(7), while the "fast"-exchanging oxazolone structure represents the remainder (70%). Intriguingly, for b(8) only the macrocycle structure is identified, which is also consistent with the "slow" kinetic rate in the HDX results. In a control experiment, a protonated cyclic peptide with 6 amino acid residues, cyclo(Gln-Trp-Phe-Gly-Leu-Met), is confirmed not to adopt an oxazolone structure, even upon collisional activation. These results demonstrate that in some cases larger macrocycle structures are surprisingly stable. While more studies are required to establish the general propensity for cyclization in b fragments, the implications from this study are troubling in terms of faulty sequence identification.

URL<Go to ISI>://000273615800038
Division

GUTHZ

Department

Molecular Dynamics

PID

98612d78c3a7bdcc65650921b997a0a2

Alternate TitleJ. Am. Chem. Soc.

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