LC/MS/MS Analysis of N-Terminal Protein Adducts with Improved Sensitivity: A Comparison of Selected Edman Isothiocyanate Reagents
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 compound...
Saved in:
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2009-01-01
|
| Series: | International Journal of Analytical Chemistry |
| Online Access: | http://dx.doi.org/10.1155/2009/153472 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832550075863138304 |
|---|---|
| author | Per Rydberg Hans von Stedingk Jörgen Magnér Jonas Björklund |
| author_facet | Per Rydberg Hans von Stedingk Jörgen Magnér Jonas Björklund |
| author_sort | Per Rydberg |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-6d364f2ddb01445c8c51cef5b7d7fd02 |
| institution | Kabale University |
| issn | 1687-8760 1687-8779 |
| language | English |
| publishDate | 2009-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | International Journal of Analytical Chemistry |
| spelling | doaj-art-6d364f2ddb01445c8c51cef5b7d7fd022025-02-03T06:07:48ZengWileyInternational Journal of Analytical Chemistry1687-87601687-87792009-01-01200910.1155/2009/153472153472LC/MS/MS Analysis of N-Terminal Protein Adducts with Improved Sensitivity: A Comparison of Selected Edman Isothiocyanate ReagentsPer Rydberg0Hans von Stedingk1Jörgen Magnér2Jonas Björklund3Arrhenius Laboratory, Department of Environmental Chemistry, Stockholm University, 106 91 Stockholm, SwedenArrhenius Laboratory, Department of Environmental Chemistry, Stockholm University, 106 91 Stockholm, SwedenDepartment of Applied Environmental Science, Stockholm University, 106 91 Stockholm, SwedenPerkinElmer Sverige AB, Oxfordhuset, Johanneslundsvägen 2, 194 81 Upplands Väsby, SwedenThis 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.http://dx.doi.org/10.1155/2009/153472 |
| spellingShingle | Per Rydberg Hans von Stedingk Jörgen Magnér Jonas Björklund LC/MS/MS Analysis of N-Terminal Protein Adducts with Improved Sensitivity: A Comparison of Selected Edman Isothiocyanate Reagents International Journal of Analytical Chemistry |
| title | LC/MS/MS Analysis of N-Terminal Protein Adducts with Improved Sensitivity: A Comparison of Selected Edman Isothiocyanate Reagents |
| title_full | LC/MS/MS Analysis of N-Terminal Protein Adducts with Improved Sensitivity: A Comparison of Selected Edman Isothiocyanate Reagents |
| title_fullStr | LC/MS/MS Analysis of N-Terminal Protein Adducts with Improved Sensitivity: A Comparison of Selected Edman Isothiocyanate Reagents |
| title_full_unstemmed | LC/MS/MS Analysis of N-Terminal Protein Adducts with Improved Sensitivity: A Comparison of Selected Edman Isothiocyanate Reagents |
| title_short | LC/MS/MS Analysis of N-Terminal Protein Adducts with Improved Sensitivity: A Comparison of Selected Edman Isothiocyanate Reagents |
| title_sort | lc ms ms analysis of n terminal protein adducts with improved sensitivity a comparison of selected edman isothiocyanate reagents |
| url | http://dx.doi.org/10.1155/2009/153472 |
| work_keys_str_mv | AT perrydberg lcmsmsanalysisofnterminalproteinadductswithimprovedsensitivityacomparisonofselectededmanisothiocyanatereagents AT hansvonstedingk lcmsmsanalysisofnterminalproteinadductswithimprovedsensitivityacomparisonofselectededmanisothiocyanatereagents AT jorgenmagner lcmsmsanalysisofnterminalproteinadductswithimprovedsensitivityacomparisonofselectededmanisothiocyanatereagents AT jonasbjorklund lcmsmsanalysisofnterminalproteinadductswithimprovedsensitivityacomparisonofselectededmanisothiocyanatereagents |