Electron transfer dissociation of doubly charged ions with different cationizing agents

Electron transfer dissociation (ETD) studies have been performed on a peptide and a synthetic polysaccharide doubly charged by different cationization agents. The ETD of protonated-sodiated bombesin gave rise to contiguous series of abundant c- and z-type ions that identified the complete sequence. ETD of the doubly protonated peptide produced a different fragment distribution, which also allowed for complete sequence coverage, but the relative intensities of some sequence ions were very small. Collisionally activated dissociation (CAD) of either precursor rendered limited sequence information. ETD of the sodiated-ammoniated pentamer of a starch-derived linear polysaccharide caused extensive fragmentation through cross-ring cleavages that revealed the possible position of a hydroxyethyl substituent on the saccharide ring. In contrast, ETD of the di-sodiated pentasaccharide did not produce a structure-revealing fragmentation pattern. On the other hand, CAD resulted in efficient glycosidic bond cleavages, either directly (from the sodiated-ammoniated precursor) or via multi-stage fragmentation (from the di-sodiated precursor), which indicated that hydroxyethylation occurs randomly at any saccharide repeat unit along the chain. Overall, the use of different cationizing agents complements the information available by using identical charge sites and opens or enhances ETD pathways that unveil valuable sequence or positional information.