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Doubly Protonated Species Collision Induced Dissociation for Identification of Isocyclosporins

preprint
submitted on 04.09.2020 and posted on 07.09.2020 by Anna Baranova, Alexey Chistov, Maxim Shuvalov, Anton Tyurin, Mikhail Biryukov, Igor Ivanov, Vera Sadykova, Aleksander Kurakov, Albina Sregeeva, Vladimir Korshun, Vera Alferova
Nonribosomal cyclopeptide cyclosporin A (CsA), produced by fungus Tolypocladium inflatum, is an extremely important immunosuppressive drug used in organ transplantations and for therapy of autoimmune diseases. Here we report for the first time production of CsA, along with related cyclosporins B and C, by Tolypocladium inflatum strains of marine origin (White Sea). Cyclosporins A–C contain an unusual amino acid, (4R)-4-((E)-2-butenyl)-4,N-dimethyl-l-threonine (MeBmt), and are prone to isomerization to non-active isocyclosporine by N→O acyl shift of valine connected to MeBmt in acidic conditions. CsA and isoCsA are not distinguishable in MS analysis of [M+H]+ ions due to the rapid [CsA+H]+→[isoCsA+H]+ conversion. We found that the N→O acyl shift is completely suppressed in cyclosporine [M+2H]2+ ions, and their MS/MS fragmentation can be used for rapid and unambiguous analysis of cyclosporins and isocylosporins. The fragmentation patterns of [CyA+2H]2+ and [isoCyA+2H]2+ ions were analyzed and explained. The developed approach could be useful for MS analysis of other peptides containing β-hydroxy-α-amino acids.

Funding

RFBR 20-33-70215 and 18-29-25073

History

Email Address of Submitting Author

v-korshun@yandex.ru

Institution

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry

Country

Russia

ORCID For Submitting Author

0000-0001-9436-6561

Declaration of Conflict of Interest

none

Version Notes

version 01

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