CZE of Sulfur-Containing Amino Acids and Peptides and Its Application to the Quantitative Study of Heavy Metal-Caused Thiol Oxidations

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• 作者：Anne-Christine Schmidt (1) Anne-Christine.Schmidt@chemie.tu-freiberg.de
  Stefanie Lauckner (1)
  Katja Lindner (1)
• 关键词：Capillary zone electrophoresis &#8211 ; Cysteine &#8211 ; Glutathione &#8211 ; Thiol oxidation &#8211 ; Heavy metals
• 刊名：Chromatographia
• 出版年：2012
• 出版时间：June 2012
• 年：2012
• 卷：75
• 期：11-12
• 页码：661-670
• 全文大小：1.1 MB
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• 作者单位：1. Institute of Analytical Chemistry, Technical University Bergakademie Freiberg, Leipziger Stra脽e 29, 09599 Freiberg, Germany
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Analytical Chemistry
  Organic Chemistry
  Pharmacy
  Biochemistry
  Plant Sciences
  Measurement Science and Instrumentation
  
• 出版者：Vieweg Verlag
• ISSN：1612-1112

文摘

The redox-active, sulfur-containing amino acids cysteine and methionine, the tripeptide glutathione and their oxidized counterparts cystine, methionine sulfoxide, and glutathione disulfide were separated as anions by capillary zone electrophoresis (CZE) in a 72 cm long fused silica capillary filled with 100 mM phosphate buffer, pH 8.0, at a voltage of +30 kV in 20 min. The optimized CZE method was suited for the implementation of quantitative metal interaction studies of the biomolecules in a biologically relevant concentration range (μM–mM). Decreasing peak areas of the reduced forms of cysteine and glutathione and simultaneously increasing peak areas of the oxidized forms after incubation of the reduced biomolecules with divalent heavy metal cations indicated redox reactions which could be responsible for toxic metal actions in biological systems. CZE measurements revealed that a 50 % oxidation grade of cysteine was achieved at a molar metal:cysteine ratio of 0.85 in case of Zn(II) addition and of 0.11 in case of Cu(II) addition, respectively. Cu(II) oxidized 50 % of the initial glutathione at a molar Cu:peptide ratio of 0.036, whereas the 50 % oxidation grade was not reached after incubation with Co(II) up to a molar ratio of Co:peptide of 0.25.