Artikel: Isotope-coded affinity tag (ICAT) approach to redox proteomics: identification and quantitation of oxidant-sensitive cysteine thiols in complex protein mixtures.
2004 Band 3, Heft 6, Seite(n) 1228–1233
Abstract: An approach is described for the simultaneous identification and quantitation of oxidant-sensitive cysteine thiols in a complex protein mixture using a thiol-specific, acid-cleavable isotope-coded affinity tag (ICAT) reagent (Applied Biosystems, USA). ... ...
Abstract | An approach is described for the simultaneous identification and quantitation of oxidant-sensitive cysteine thiols in a complex protein mixture using a thiol-specific, acid-cleavable isotope-coded affinity tag (ICAT) reagent (Applied Biosystems, USA). The approach is based on the fact that only free cysteine thiols are susceptible to labeling by the iodoacetamide-based ICAT, and that mass spectrometry can be used to quantitate the relative labeling of free thiols. Applying this approach, we have identified cysteine thiols of proteins in a rabbit heart membrane fraction that are sensitive to a high concentration of hydrogen peroxide. Previously known and some novel proteins with oxidant-sensitive cysteines were identified. Of the many protein thiols labeled by the ICAT, only relatively few were oxidized more than 50% despite the high concentration of oxidant used, indicating that oxidant-sensitive thiols are relatively rare, and denoting their specificity and potential functional relevance. |
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Mesh-Begriff(e) | Affinity Labels ; Animals ; Carbon Isotopes ; Cysteine/analysis ; Cysteine/metabolism ; Hydrogen Peroxide/metabolism ; Membrane Proteins/analysis ; Myocardium/chemistry ; Oxidants/metabolism ; Oxidation-Reduction ; Proteins/analysis ; Proteins/metabolism ; Proteomics/methods ; Rabbits ; Sulfhydryl Compounds/analysis ; Sulfhydryl Compounds/metabolism |
Chemische Substanzen | Affinity Labels ; Carbon Isotopes ; Membrane Proteins ; Oxidants ; Proteins ; Sulfhydryl Compounds ; Hydrogen Peroxide (BBX060AN9V) ; Cysteine (K848JZ4886) |
Sprache | Englisch |
Erscheinungsdatum | 2004-11 |
Erscheinungsland | United States |
Dokumenttyp | Journal Article ; Research Support, U.S. Gov't, P.H.S. |
ZDB-ID | 2078618-9 |
ISSN | 1535-3893 |
ISSN | 1535-3893 |
DOI | 10.1021/pr049887e |
Datenquelle | MEDical Literature Analysis and Retrieval System OnLINE |
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