Can dimedone be used to study selenoproteins? An investigation into the reactivity of dimedone toward oxidized forms of selenocysteine.

Autor: Payne NC; Department of Biochemistry, University of Vermont, College of Medicine, Burlington, Vermont, 05405., Barber DR; Department of Biochemistry, University of Vermont, College of Medicine, Burlington, Vermont, 05405., Ruggles EL; Department of Biochemistry, University of Vermont, College of Medicine, Burlington, Vermont, 05405., Hondal RJ; Department of Biochemistry, University of Vermont, College of Medicine, Burlington, Vermont, 05405.
Jazyk: angličtina
Zdroj: Protein science : a publication of the Protein Society [Protein Sci] 2019 Jan; Vol. 28 (1), pp. 41-55. Date of Electronic Publication: 2018 Mar 10.
DOI: 10.1002/pro.3390
Abstrakt: Dimedone is a widely used reagent to assess the redox state of cysteine-containing proteins as it will alkylate sulfenic acid residues, but not sulfinic acid residues. While it has been reported that dimedone can label selenenic acid residues in selenoproteins, we investigated the stability, and reversibility of this label in a model peptide system. We also wondered whether dimedone could be used to detect seleninic acid residues. We used benzenesulfinic acid, benzeneseleninic acid, and model selenocysteine-containing peptides to investigate possible reactions with dimedone. These peptides were incubated with H 2 O 2 in the presence of dimedone and then the reactions were followed by liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS). The native peptide, H-PTVTGCUG-OH (corresponding to the native amino acid sequence of the C-terminus of mammalian thioredoxin reductase), could not be alkylated by dimedone, but could be carboxymethylated with iodoacetic acid. However the "mutant peptide," H-PTVTGAUG-OH, could be labeled with dimedone at low concentrations of H 2 O 2 , but the reaction was reversible by addition of thiol. Due to the reversible nature of this alkylation, we conclude that dimedone is not a good reagent for detecting selenenic acids in selenoproteins. At high concentrations of H 2 O 2 , selenium was eliminated from the peptide and a dimeric form of dimedone could be detected using LCMS and 1 H NMR. The dimeric dimedone product forms as a result of a seleno-Pummerer reaction with Sec-seleninic acid. Overall our results show that the reaction of dimedone with oxidized cysteine residues is quite different from the same reaction with oxidized selenocysteine residues.
(© 2018 The Protein Society.)
Databáze: MEDLINE
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