Simultaneous determination of nitrated and oligomerized proteins by size exclusion high-performance liquid chromatography coupled to photodiode array detection
- Chemical modifications such as nitration and cross-linking may enhance the allergenic potential of proteins. The kinetics and mechanisms of the underlying chemical processes, however, are not yet well understood. Here, we present a size-exclusion chromatography/spectrophotometry method (SEC-HPLC-DAD) that allows a simultaneous detection of mono-, di-, tri-, and higher protein oligomers, as well as their individual nitration degrees (NDs). The ND results of proteins from this new method agree well with the results from an alternative well-established method, for the analysis of tetranitromethane (TNM)- and nitrogen dioxide and ozone (NO2/O3)-nitrated protein samples. Importantly, the NDs for individual oligomer fractions can be obtained from the new method, and also, we provide a proof of principle for the calculation of the concentrations for individual protein oligomer fractions by their determined NDs, which will facilitate the investigation of the kinetics and mechanism for proteinChemical modifications such as nitration and cross-linking may enhance the allergenic potential of proteins. The kinetics and mechanisms of the underlying chemical processes, however, are not yet well understood. Here, we present a size-exclusion chromatography/spectrophotometry method (SEC-HPLC-DAD) that allows a simultaneous detection of mono-, di-, tri-, and higher protein oligomers, as well as their individual nitration degrees (NDs). The ND results of proteins from this new method agree well with the results from an alternative well-established method, for the analysis of tetranitromethane (TNM)- and nitrogen dioxide and ozone (NO2/O3)-nitrated protein samples. Importantly, the NDs for individual oligomer fractions can be obtained from the new method, and also, we provide a proof of principle for the calculation of the concentrations for individual protein oligomer fractions by their determined NDs, which will facilitate the investigation of the kinetics and mechanism for protein tyrosine nitration and cross-linking.…
Autor*innen: | F. Liu, K. Reinmuth-Selzle, S. Lai, Michael G. WellerORCiD, U. Pöschl, C. J. Kampf |
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Dokumenttyp: | Zeitschriftenartikel |
Veröffentlichungsform: | Verlagsliteratur |
Sprache: | Englisch |
Titel des übergeordneten Werkes (Englisch): | Journal of Chromatography A |
Jahr der Erstveröffentlichung: | 2017 |
Verlag: | Elsevier B.V. |
Verlagsort: | Amsterdam |
Jahrgang/Band: | 1495 |
Erste Seite: | 76 |
Letzte Seite: | 82 |
DDC-Klassifikation: | Naturwissenschaften und Mathematik / Chemie / Analytische Chemie |
Technik, Medizin, angewandte Wissenschaften / Ingenieurwissenschaften / Sanitär- und Kommunaltechnik; Umwelttechnik | |
Freie Schlagwörter: | Air pollution; Albumin; BSA; Dimer; HPLC-DAD; Nitrogen oxides; Nitrotyrosine; Oligomer; Protein; Protein nitration degree; Protein oligomer analysis; Size exclusion chromatography; Tetranitromethane; Trimer |
DOI: | 10.1016/j.chroma.2017.03.015 |
URL: | http://www.sciencedirect.com/science/article/pii/S0021967317303795 |
ISSN: | 0021-9673 |
Verfügbarkeit des Dokuments: | Datei im Netzwerk der BAM verfügbar ("Closed Access") |
Datum der Freischaltung: | 22.05.2017 |
Referierte Publikation: | Ja |
Datum der Eintragung als referierte Publikation: | 22.05.2017 |