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LCMSMS Analysis of N-Terminal Protein Adducts with Improved Sensitivity : A Comparison of Selected Edman Isothiocyanate Reagents
Joint Authors
Magnér, Jörgen
Rydberg, Per
von Stedingk, Hans
Björklund, Jonas
Source
International Journal of Analytical Chemistry
Issue
Vol. 2009, Issue 2009 (31 Dec. 2009), pp.1-13, 13 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2009-11-04
Country of Publication
Egypt
No. of Pages
13
Main Subjects
Abstract EN
This study provides a basis for a new and straightforward method for LC/MS/MS-based screening of N-terminal protein adducts.
This procedure is denoted the “FIRE procedure” as fluorescein isothiocyanate (FITC) gave superior sensitivity by LC/MS/MS when measuring adducts (R) of electrophilic compounds with a modified Edman procedure.
The principles of the FIRE-procedure are that adducts to N-terminal amino acids selectively are detached and measured from of proteins after derivatisation by isothiocyanate Edman reagents.
In this study, FITC, 4-N,N-dimethylaminoazobenzene 4′-isothiocyanate (DABITC) and 4-dimethylamino-1-naphthyl isothiocyanate (DNITC) were used to synthesize thiohydantoin analytes from valine and N-methylvaline.
The sensitivity by LC/MS/MS was enhanced by up to three orders of magnitude as compared to phenyl isothiocyanate and higher as compared to pentafluorophenyl isothiocyanate.
The FITC reagent will enable measurements of low background adduct levels.
Synthesized analytes were characterised with, for example, H1 NMR, C13 NMR, LC/MS/MS, and UV.
American Psychological Association (APA)
Rydberg, Per& von Stedingk, Hans& Magnér, Jörgen& Björklund, Jonas. 2009. LCMSMS Analysis of N-Terminal Protein Adducts with Improved Sensitivity : A Comparison of Selected Edman Isothiocyanate Reagents. International Journal of Analytical Chemistry،Vol. 2009, no. 2009, pp.1-13.
https://search.emarefa.net/detail/BIM-450056
Modern Language Association (MLA)
Rydberg, Per…[et al.]. LCMSMS Analysis of N-Terminal Protein Adducts with Improved Sensitivity : A Comparison of Selected Edman Isothiocyanate Reagents. International Journal of Analytical Chemistry No. 2009 (2009), pp.1-13.
https://search.emarefa.net/detail/BIM-450056
American Medical Association (AMA)
Rydberg, Per& von Stedingk, Hans& Magnér, Jörgen& Björklund, Jonas. LCMSMS Analysis of N-Terminal Protein Adducts with Improved Sensitivity : A Comparison of Selected Edman Isothiocyanate Reagents. International Journal of Analytical Chemistry. 2009. Vol. 2009, no. 2009, pp.1-13.
https://search.emarefa.net/detail/BIM-450056
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-450056