Differential fluorescence labeling of cysteinyl clusters uncovers high tissue levels of thionein

Proceedings of the National Academy of Sciences of the United States of America
Y YangB L Vallee

Abstract

The isolation of thionein (T) from tissues has not been reported heretofore. T contains 20 cysteinyl residues that react with 7-fluorobenz-2-oxa-1,3-diazole-4-sulfonamide to form fluorescent adducts. In metallothionein (MT) the cysteinyl residues, which are bound to zinc, do not react. However, they do react in the presence of a chelating agent such as EDTA. The resultant difference in chemical reactivity provides a means to measure T in the absence of EDTA, (MT + T) in its presence, and, of course, MT by difference. The 7-fluorobenz-2-oxa-1,3-diazole-4-sulfonamide derivative of T can be isolated from tissue homogenates by HPLC and quantified fluorimetrically with a detection limit in the femtomolar range and a linear response over 3 orders of magnitude. Analysis of liver, kidney, and brain of rats reveals almost as much T as MT. Moreover, in contrast to earlier views, MT in tissue extracts appears to be less stable than T. The existence of T in tissues under normal physiological conditions has important implications for its function both in zinc metabolism and the redox balance of the cell.

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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
Mar 30, 2005·Toxicology Letters·John H BeattieHeather M Wallace
Sep 5, 2003·Chemistry & Biology·Niroshini M GilesClaus Jacob
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Mar 12, 2010·Chemical Communications : Chem Comm·Mercè CapdevilaEsteve Fàbregas
Sep 22, 2006·Antioxidants & Redox Signaling·Wolfgang Maret
Sep 22, 2006·Antioxidants & Redox Signaling·Irina Korichneva
Sep 21, 2010·Antioxidants & Redox Signaling·Mandar A Aras, Elias Aizenman
Jul 12, 2007·Molecular Medicine·Wolfgang Maret, Artur Krezel
Jan 21, 2014·Progress in Neurobiology·Ivo F ScheiberRalf Dringen
Oct 25, 2001·Proceedings of the National Academy of Sciences of the United States of America·W Maret
Jun 1, 2006·Proceedings of the National Academy of Sciences of the United States of America·Daphna SagherHerbert Weissbach
Mar 6, 2003·Proceedings of the National Academy of Sciences of the United States of America·Sung-Hye Hong, Wolfgang Maret
Jan 9, 2009·Biometals : an International Journal on the Role of Metal Ions in Biology, Biochemistry, and Medicine·Wolfgang Maret
Jun 16, 2010·Biometals : an International Journal on the Role of Metal Ions in Biology, Biochemistry, and Medicine·Ivan SabolićCarol M Herak-Kramberger
Aug 7, 2013·Biometals : an International Journal on the Role of Metal Ions in Biology, Biochemistry, and Medicine·Roobee GarlaMohan L Garg
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Jul 8, 2016·International Journal of Molecular Sciences·Francisco Andrés Peralta, Juan Pablo Huidobro-Toro
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Dec 13, 2007·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Artur Krezel, Wolfgang Maret
Oct 11, 2008·Electrophoresis·Hajo Haase, Wolfgang Maret

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