A differential assay for the reduced and oxidized states of metallothionein and thionein

Analytical Biochemistry
Hajo Haase, Wolfgang Maret

Abstract

In the cellular environment, the sulfur ligands in zinc/thiolate coordination sites of proteins can be oxidized with concomitant mobilization of zinc. The characterization of such "redox zinc switches" requires the determination of three species, i.e., the zinc-containing complex and the zinc-free complex with the thiolate ligands either reduced or oxidized. Differential chemical modification of thiol groups in the presence and absence of either reducing or chelating agents allows the analytical speciation of such systems as demonstrated here for the characterization of the redox and metal-binding states of mammalian metallothionein. Thiol derivatization with 6-iodoacetamidofluorescein in the presence and absence of the reducing agent tris(2-carboxyethyl)phosphine, high-performance liquid chromatographic separation, and photometric detection are employed to determine the reduced and oxidized protein. Because the holoprotein reacts only in the presence of a chelating agent such as ethylenediaminetetraacetate (EDTA) its amount can be determined as the difference between measurements in the presence and the absence of EDTA. This method is applied to the study of the chemical and enzymatic oxidation of metallothionein/thionein. It ...Continue Reading

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Citations

Sep 15, 2005·Biometals : an International Journal on the Role of Metal Ions in Biology, Biochemistry, and Medicine·Hajo Haase, Wolfgang Maret
Jun 1, 2013·Journal of Toxicology and Environmental Health. Part a·F GagnéC Gagnon
Sep 14, 2013·Journal of Toxicology and Environmental Health. Part a·F GagnéC Gagnon
Sep 22, 2006·Antioxidants & Redox Signaling·Wolfgang Maret
Apr 30, 2009·Annual Review of Nutrition·Hajo Haase, Lothar Rink
Jul 12, 2007·Molecular Medicine·Wolfgang Maret, Artur Krezel
Apr 5, 2014·Aquatic Toxicology·Phenny MwaangaPeter van den Hurk
Jun 8, 2011·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Wolfgang Maret
Sep 14, 2010·Biochemical and Biophysical Research Communications·Thanh T NguMartin J Stillman
Apr 20, 2010·Biochemical and Biophysical Research Communications·Thanh T NguMartin J Stillman
Jan 28, 2009·Biochemical and Biophysical Research Communications·Gerd WellenreutherHajo Haase
Oct 26, 2005·Journal of Trace Elements in Medicine and Biology : Organ of the Society for Minerals and Trace Elements (GMS)·Wolfgang Maret
Nov 24, 2004·Biochemical and Biophysical Research Communications·Kelly E Rigby, Martin J Stillman
May 24, 2016·Comparative Biochemistry and Physiology. Toxicology & Pharmacology : CBP·F GagnéC Gagnon
Mar 30, 2007·Archives of Biochemistry and Biophysics·Artur KrezelWolfgang Maret
Jul 3, 2018·Biological & Pharmaceutical Bulletin·Shinsuke NakamuraHideaki Hara
Oct 11, 2008·Electrophoresis·Hajo Haase, Wolfgang Maret
Oct 11, 2005·American Journal of Physiology. Heart and Circulatory Physiology·Murat Ayaz, Belma Turan
May 25, 2005·Protein Engineering, Design & Selection : PEDS·S-H HongW Maret
Dec 1, 2006·The Biochemical Journal·Artur Krezel, Wolfgang Maret
Mar 29, 2020·Metallomics : Integrated Biometal Science·Amelia T YuanMartin J Stillman
Mar 5, 2021·Redox Biology·Christopher Hübner, Hajo Haase
Dec 5, 2008·Journal of the American Chemical Society·Thanh T NguMartin J Stillman
Jul 11, 2009·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Wolfgang Maret
Oct 16, 2021·Chemical Reviews·Artur Krężel, Wolfgang Maret

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