Spontaneous N --> O acyl shift in the [M + H](+) ions of [MeBmt1]-cyclosporins in an ion trap.

In an ion trap the protonated molecules of the cyclic undecapeptides cyclosporins having 3-hydroxy-4-methyl-2-methylamino-6-octenoic acid (MeBmt) in their backbone undergo an N --> O peptidyl shift into the corresponding [M + H](+) ions of isocyclosporins. This rearrangement does not take place in cyclosporins [Bmt1]Cs and [3'-deoxy-MeBmt1]Cs. In cyclosporin [Thr2]Cs having two threonines in the molecule, only one of them participates in the N,O-acyl transfer. It can be concluded that the presence of the basic N-methylamino group of MeBmt, which can serve as the primary site of protonation, is necessary for isocyclosporin formation. A dominating ion series originating from the primary cleavage between MeBmt (first position in the cyclosporin ring) and amino acid residue at the neighbouring eleventh position is then observed in collision-induced dissociation spectra of protonated molecules of cyclosporins. This 'isocyclosporin' ion series can effectively be used for easy and complete cyclosporin sequencing using a tandem mass spectrometric (MS3) experiment in an ion trap. The paper further introduces an improved Gross mass spectral nomenclature for cyclic peptide sequencing and several techniques for the generation of protonated molecules of cyclosporins. Their preparation represents the fundamental requirement for smooth sequencing of cyclosporins by tandem mass spectral techniques.