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Charge site manipulation to enhance top‐down fragmentation efficiency

Habeck, Tanja ; Maciel, Edvaldo Vasconcelos Soares ; Kretschmer, Kevin ; Lermyte, Frederik (2024)
Charge site manipulation to enhance top‐down fragmentation efficiency.
In: Proteomics : Proteomics and Systems Biology, 24 (3-4)
doi: 10.1002/pmic.202300082
Artikel, Bibliographie

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Kurzbeschreibung (Abstract)

In recent years, top‐down mass spectrometry has become a widely used approach to study proteoforms; however, improving sequence coverage remains an important goal. Here, two different proteins, α‐synuclein and bovine carbonic anhydrase, were subjected to top‐down collision‐induced dissociation (CID) after electrospray ionisation. Two high‐boiling solvents, DMSO and propylene carbonate, were added to the protein solution in low concentration (2%) and the effects on the top‐down fragmentation patterns of the proteins were systematically investigated. Each sample was measured in triplicate, which revealed highly reproducible differences in the top‐down CID fragmentation patterns in the presence of a solution additive, even if the same precursor charge state was isolated in the quadrupole of the instrument. Further investigation supports the solution condition‐dependent selective formation of different protonation site isomers as the underlying cause of these differences. Higher sequence coverage was often observed in the presence of additives, and the benefits of this approach became even more evident when datasets from different solution conditions were combined, as increases up to 35% in cleavage coverage were obtained. Overall, this approach therefore represents a promising opportunity to increase top‐down fragmentation efficiency.

Typ des Eintrags: Artikel
Erschienen: 2024
Autor(en): Habeck, Tanja ; Maciel, Edvaldo Vasconcelos Soares ; Kretschmer, Kevin ; Lermyte, Frederik
Art des Eintrags: Bibliographie
Titel: Charge site manipulation to enhance top‐down fragmentation efficiency
Sprache: Englisch
Publikationsjahr: Februar 2024
Ort: Weinheim
Verlag: Wiley-VCH
Titel der Zeitschrift, Zeitung oder Schriftenreihe: Proteomics : Proteomics and Systems Biology
Jahrgang/Volume einer Zeitschrift: 24
(Heft-)Nummer: 3-4
Kollation: 10 Seiten
DOI: 10.1002/pmic.202300082
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Kurzbeschreibung (Abstract):

In recent years, top‐down mass spectrometry has become a widely used approach to study proteoforms; however, improving sequence coverage remains an important goal. Here, two different proteins, α‐synuclein and bovine carbonic anhydrase, were subjected to top‐down collision‐induced dissociation (CID) after electrospray ionisation. Two high‐boiling solvents, DMSO and propylene carbonate, were added to the protein solution in low concentration (2%) and the effects on the top‐down fragmentation patterns of the proteins were systematically investigated. Each sample was measured in triplicate, which revealed highly reproducible differences in the top‐down CID fragmentation patterns in the presence of a solution additive, even if the same precursor charge state was isolated in the quadrupole of the instrument. Further investigation supports the solution condition‐dependent selective formation of different protonation site isomers as the underlying cause of these differences. Higher sequence coverage was often observed in the presence of additives, and the benefits of this approach became even more evident when datasets from different solution conditions were combined, as increases up to 35% in cleavage coverage were obtained. Overall, this approach therefore represents a promising opportunity to increase top‐down fragmentation efficiency.

Freie Schlagworte: electrospray ionization mass spectrometry (ESI‐MS), mass spectrometry, top‐down proteomics
ID-Nummer: Artikel-ID: 2300082
Zusätzliche Informationen:

Special Issue: Top‐down proteomics and proteoforms

Sachgruppe der Dewey Dezimalklassifikatin (DDC): 500 Naturwissenschaften und Mathematik > 540 Chemie
500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie
Fachbereich(e)/-gebiet(e): 07 Fachbereich Chemie
07 Fachbereich Chemie > Clemens-Schöpf-Institut > Fachgebiet Biochemie
07 Fachbereich Chemie > Clemens-Schöpf-Institut
Hinterlegungsdatum: 18 Jun 2024 05:43
Letzte Änderung: 18 Jun 2024 08:39
PPN: 519212665
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