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Titel
Topdown and bottomup characterization of nitrated birch pollen allergen Bet v 1a with CZE hyphenated to an Orbitrap mass spectrometer
VerfasserGusenkov, Sergey ; Stutz, Hanno
Erschienen in
Electrophoresis, Hoboken, 2018, Jg. 39, H. 9-10, S. 1190-1200
ErschienenHoboken : Wiley, 2018
SpracheEnglisch
DokumenttypAufsatz in einer Zeitschrift
Schlagwörter (EN)Bottomup / CZEESIOrbitrap MS / Laboratorybuilt ESI device / Nitration sites of pollen allergen / Topdown
ISSN1522-2683
URNurn:nbn:at:at-ubs:3-11582 Persistent Identifier (URN)
DOI10.1002/elps.201700413 
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Topdown and bottomup characterization of nitrated birch pollen allergen Bet v 1a with CZE hyphenated to an Orbitrap mass spectrometer [1.57 mb]
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Zusammenfassung (Englisch)

Tyrosine (Tyr) residues of the major pollen allergen of birch Betula verrucosa, Bet v 1a, were nitrated by peroxynitrite. This modification enhances the allergenicity. Modified tyrosines were identified by analyzing intact allergen variants in combination with topdown and bottomup approaches. Therefore, a laboratorybuilt sheathliquid assisted ESI interface was applied for hyphenation of CE to an Orbitrap mass spectrometer to localize individual nitration sites. The major focus was on identification of primary nitration sites. The topdown approach unambiguously identified Tyr 5 as the most prominent modification site. Fragments from the allergen core and the Cterminal part carried up to three potential nitration sites, respectively. Thus, a bottomup approach with tryptic digest was used as a complementary strategy which allowed for the unambiguous localization of nitration sites within the respective peptides. Nitration propensity for individual Tyr residues was addressed by comparison of MS signals of nitrated peptides relative to all cognates of homolog primary sequence. Combined data identified surface exposed Tyr 5 and Tyr 66 as major nitration sites followed by less accessible Tyr 158 whereas Tyr 81, 83 and 150 possess a lower nitration tendency and are apparently modified in variants with higher nitration levels.

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