Broadband detection electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry to reveal enzymatically and chemically induced deamidation reactions within peptides
| Autor: | Florian von der Mülbe, Dietmar G. Schmid, Günther Jung, and Toni Weinschenk, Burkhard Fleckenstein |
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| Rok vydání: | 2002 |
| Předmět: |
Spectrometry
Mass Electrospray Ionization Protein mass spectrometry Electrospray ionization Guinea Pigs Top-down proteomics Mass spectrometry Fourier transform ion cyclotron resonance Gliadin Analytical Chemistry Epitopes Animals Humans Deamidation B-Lymphocytes Chromatography Transglutaminases Fourier Analysis Chemistry Hydrolysis Selected reaction monitoring Cyclotrons Amides Peptide Fragments Celiac Disease Monoisotopic mass Peptides |
| Zdroj: | Analytical chemistry. 73(24) |
| ISSN: | 0003-2700 |
| Popis: | Among the numerous forms of chemical degradation of peptides or proteins, deamidation is one of the alterations observed most frequently. In this irreversible reaction, a glutamine or an asparagine side chain is hydrolyzed to glutamic acid or aspartic acid, respectively (conversion of NH2 to OH). Besides its influence in the deterioration of biotechnological and food products, deamidation represents a defined posttranslational modification reaction with respect to proteomics. Here mass spectrometric techniques play a leading role in determining posttranslational modifications. However, not all mass spectrometers are able to resolve signal differences of 0.0193 Da (mass difference of 12CO vs 13CNH) for singly charged molecules, the mass difference between the first isotopic signal of an asparagine/glutamine-containing peptide and the monoisotopic signal of the corresponding partially deamidated aspartate/glutamate derivative. To detect partial deamidation within peptides, advantage has been taken of the ability of Fourier transform ion cyclotron resonance mass spectrometry to perform very high mass resolution. In this work, we investigated up to triply charged ions produced by electrospray ionization using direct infusion. Although the special heterodyne detection mode enables higher mass resolution than the routinely used broadband detection, often only a small mass window can be investigated. Using broadband detection, we were able to resolve ions with a difference of m/z 0.0064 to detect partially deamidated peptides formed either enzymatically or under acidic and basic conditions. |
| Databáze: | OpenAIRE |
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