The role of lysine ɛ-amine group on the macrocyclization of b ions

A study was carried out to examine if the amine (NH2) group located on the side chains of lysine (K), glutamine (Q), or asparagine (N) residue has any effect on the macrocyclization of b ions even though the N-terminals of the peptides were acetylated. The work utilized the model peptides Ac-KYAGFLVG, Ac-QYAGFLV-NH2, and Ac-NYAGFLV-NH2. The CID mass spectra of b7 ions originated from these three peptides exhibited that the macrocyclization still occurred for the lysine containing peptide in spite of the N-terminal of the peptide was acetylated, but was failed to be observed for glutamine and asparagine containing peptides. These current results reveal that the lysine side chain ɛ-amine group has been involved in the macrocyclization of the peptide b ions for the N-terminal acetylated peptides and consequently, non-direct sequence b ions were observed in the CID mass spectra. However, due to the amide group on the side chains of the glutamine and asparagine residues, the nucleophilicity of their groups greatly reduced; therefore the scrambling b ions were not detected in their b7 ion CID mass spectra. In addition, the effect of the lysine position was also studied for series of six isomeric octapeptides such as, Ac-KYAGFLVG, Ac-YKAGFLVG, Ac-YAKGFLVG, Ac-YAGKFLVG, Ac-YAGFKLVG and Ac-YAGFLKVG in order to examine the relationship between the intensities of non-direct sequence b ions and the lysine position in the octapeptide series. The results clearly demonstrated that the most abundant non-direct sequence b ions were observed for the first position of lysine residue in the N-terminal acetylated octapeptide, however, when the lysine residue gets closer to the C-terminal position the relative intensities of the scrambled b ions were greatly decreased.

Graphical abstractFigure optionsDownload full-size imageDownload high-quality image (98 K)Download as PowerPoint slideHighlights► N-terminal acetylated model octapeptides containing lysine, glutamine or asparagine located at the N-terminal position 1 were studied to investigate the effect of ɛ-amine group on the macrocyclization of b ions. ► Lysine ɛ-amine group is involved in the b-type macrocyclization. ► Position of lysine residue in the N-terminal acetylated model octapeptides is important.