Mechanistic investigation of phosphate ester bond cleavages of glycylphosphoserinyltryptophan radical cations under low-energy collision-induced dissociation.

Under the conditions of low-energy collision-induced dissociation (CID), the canonical glycylphosphoserinyltryptophan radical cation having its radical located on the side chain of the tryptophan residue ([G(p)SW](•+)) fragments differently from its tautomer with the radical initially generated on the α-carbon atom of the glycine residue ([G(•)(p)SW](+)). The dissociation of [G(•)(p)SW](+) is dominated by the neutral loss of H3PO4 (98 Da), with backbone cleavage forming the [b2 - H](•+)/y1(+) pair as the minor products. In contrast, for [G(p)SW](•+), competitive cleavages along the peptide backbone, such as the formation of [G(p)SW - CO2](•+) and the [c2 + 2H](+)/[z(1) - H](•+) pair, significantly suppress the loss of neutral H3PO4. In this study, we used density functional theory (DFT) to examine the mechanisms for the tautomerizations of [G(•)(p)SW](+) and [G(p)SW](•+) and their dissociation pathways. Our results suggest that the dissociation reactions of these two peptide radical cations are more efficient than their tautomerizations, as supported by Rice-Ramsperger-Kassel-Marcus (RRKM) modeling. We also propose that the loss of H3PO4 from both of these two radical cationic tautomers is preferentially charge-driven, similar to the analogous dissociations of even-electron protonated peptides. The distonic radical cationic character of [G(•)(p)SW](+) results in its charge being more mobile, thereby favoring charge-driven loss of H3PO4; in contrast, radical-driven pathways are more competitive during the CID of [G(p)SW](•+).