Glutathionylspermidine synthetase (GspS) was introduced into human cells to label endogenous glutathione (GSH) with biotinyl spermine (biotin‐spm) for the identification of S‐thiolated proteins. The proteins carrying Gspm‐biotin can be detected by immunoblotting (A in figure). Enrichment by using the biotin tag enabled subsequent mass‐spectrometry‐based proteomic analysis (B) for site‐specific identification of glutathionylated sites.
Glutathionylspermidin‐Synthetase (GspS) wurde in menschliche Zellen eingebracht, um endogenes Glutathion (GSH) mit Biotinylspermin (Biotin‐spm) für die Identifizierung von S‐thiolierten Proteinen zu markieren. Die Proteine mit Gspm‐biotin können mit einem Immunblot nachgewiesen werden (A). Eine Anreicherung mithilfe des Biotin‐Markers ermöglicht eine massenspektrometrische Proteomanalyse (B) zur ortsspezifischen Identifikation von Glutathionylierungen.
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