American Chemical Society
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Effect of Peptide Fragment Size on the Propensity of Cyclization in Collision-Induced Dissociation: Oligoglycine b2−b8

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journal contribution
posted on 2009-12-30, 00:00 authored by Xian Chen, Long Yu, Jeffrey D. Steill, Jos Oomens, Nick C. Polfer
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 b2 to b8, 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 bn fragments (n = 2, 3) exclusively adopt oxazolone-type structures, whereas a mixture of oxazolone and macrocycle b fragment structures are formed for midsized bn 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 ∼30% of the b ion population for b6b7, while the “fast”-exchanging oxazolone structure represents the remainder (70%). Intriguingly, for b8 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.