Mobile Protons Versus Mobile Radicals: Gas-Phase Unimolecular Chemistry of Radical Cations of Cysteine-Containing Peptides⁎†

A combination of electrospray ionization (ESI), multistage, and high-resolution mass spectrometry experiments are used to examine the gas-phase fragmentation reactions of radical cations of cysteine containing di- and tripeptides. Two different chemical methods were used to form initial populations of radical cations in which the radical sites were located at different positions: (1) sulfur-centered cysteinyl radicals via bond homolysis of protonated S-nitrosocysteine containing peptides; and (2) α-carbon backbone-centered radicals via Siu's sequence of reactions (J. Am. Chem. Soc.2008, 130, 7862). Comparison of the fragmentation reactions of these regiospecifically generated radicals suggests that hydrogen atom transfer (HAT) between the α C–H of adjacent residues and the cysteinyl radical can occur. In addition, using accurate mass measurements, deuterium labeling, and comparison with an authentic sample, a novel loss of part of the N-terminal cysteine residue was shown to give rise to the protonated, truncated N-formyl peptide (an even-electron xn ion). DFT calculations were performed on the radical cation [GCG]·+ to examine: the relative stabilities of isomers with different radical and protonation sites; the barriers associated with radical migration between four possible radical sites, [G·CG]+, [GC·G]+, [GCG·]+, and [GC(S·)G]+; and for dissociation from these sites to yield b2-type ions.

Graphical AbstractTwo different methods were used to generate radical cations of cysteine containing di- and tripeptides. Fragmentation products of these were examined using experiment and DFT.Figure optionsDownload full-size imageDownload high-quality image (29 K)Download as PowerPoint slide