Zitieren Sie bitte immer diesen URN: urn:nbn:de:kobv:b43-398714
Release of free amino acids upon oxidation of peptides and proteins by hydroxyl radicals
- Hydroxyl radical-induced oxidation of proteins and peptides can lead to the cleavage of the peptide, leading to a release of fragments. Here, we used high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) and pre-column online ortho-phthalaldehyde (OPA) derivatization-based amino acid analysis by HPLC with diode array detection and fluorescence detection to identify and quantify free amino acids released upon oxidation of proteins and peptides by hydroxyl radicals. Bovine serum albumin (BSA), ovalbumin (OVA) as model proteins, and synthetic tripeptides (comprised of varying compositions of the amino acids Gly, Ala, Ser, and Met) were used for reactions with hydroxyl radicals, which were generated by the Fenton reaction of iron ions and hydrogen peroxide. The molar yields of free glycine, aspartic acid, asparagine, and alanine per peptide or protein varied between 4 and 55%. For protein oxidation reactions, the molar yields of Gly (∼32-55% for BSA, ∼10-21% for OVA)Hydroxyl radical-induced oxidation of proteins and peptides can lead to the cleavage of the peptide, leading to a release of fragments. Here, we used high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) and pre-column online ortho-phthalaldehyde (OPA) derivatization-based amino acid analysis by HPLC with diode array detection and fluorescence detection to identify and quantify free amino acids released upon oxidation of proteins and peptides by hydroxyl radicals. Bovine serum albumin (BSA), ovalbumin (OVA) as model proteins, and synthetic tripeptides (comprised of varying compositions of the amino acids Gly, Ala, Ser, and Met) were used for reactions with hydroxyl radicals, which were generated by the Fenton reaction of iron ions and hydrogen peroxide. The molar yields of free glycine, aspartic acid, asparagine, and alanine per peptide or protein varied between 4 and 55%. For protein oxidation reactions, the molar yields of Gly (∼32-55% for BSA, ∼10-21% for OVA) were substantially higher than those for the other identified amino acids (∼5-12% for BSA, ∼4-6% for OVA). Upon oxidation of tripeptides with Gly in C-terminal, mid-chain, or N-terminal positions, Gly was preferentially released when it was located at the C-terminal site. Overall, we observe evidence for a site-selective formation of free amino acids in the OH radical-induced oxidation of peptides and proteins, which may be due to a reaction pathway involving nitrogen-centered radicals.…
Autor*innen: | F. Liu, S. Lai, H. Tong, P. S. J. Lakey, M. Shiraiwa, Michael G. WellerORCiD, U. Pöschl, C. J. Kampf |
---|---|
Dokumenttyp: | Zeitschriftenartikel |
Veröffentlichungsform: | Verlagsliteratur |
Sprache: | Englisch |
Titel des übergeordneten Werkes (Englisch): | Analytical and Bioanalytical Chemistry |
Jahr der Erstveröffentlichung: | 2017 |
Veröffentlichende Institution: | Bundesanstalt für Materialforschung und -prüfung (BAM) |
Verlag: | Springer |
Verlagsort: | Heidelberg |
Jahrgang/Band: | 409 |
Ausgabe/Heft: | 9 |
Erste Seite: | 2411 |
Letzte Seite: | 2420 |
DDC-Klassifikation: | Naturwissenschaften und Mathematik / Chemie / Analytische Chemie |
Technik, Medizin, angewandte Wissenschaften / Ingenieurwissenschaften / Sanitär- und Kommunaltechnik; Umwelttechnik | |
Freie Schlagwörter: | AAA; Amino acid analysis; Bovine serum albumin; Degradation; Fragmentation; Hydroxyl radicals; LC-MS; Mechanism; Ortho-Phthalaldehyde; Ovalbumin; Oxidation; Peptides; Proteins; Tripeptides |
DOI: | 10.1007/s00216-017-0188-y |
URN: | urn:nbn:de:kobv:b43-398714 |
URL: | http://link.springer.com/article/10.1007%2Fs00216-017-0188-y |
ISSN: | 1618-2650 |
ISSN: | 1618-2642 |
Verfügbarkeit des Dokuments: | Datei für die Öffentlichkeit verfügbar ("Open Access") |
Lizenz (Deutsch): | Creative Commons - Namensnennung |
Datum der Freischaltung: | 18.04.2017 |
Referierte Publikation: | Ja |
Datum der Eintragung als referierte Publikation: | 18.04.2017 |
Schriftenreihen ohne Nummerierung: | Wissenschaftliche Artikel der BAM |