Quantification of Oxidized Metallothionein in Biological Material by a Cd Saturation Method

✍ Scribed by D. Klein; S. Sato; K.H. Summer

• Book ID: 102559258
• Publisher: Elsevier Science
• Year: 1994
• Tongue: English
• Weight: 387 KB
• Volume: 221
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.1994.1434

✦ Synopsis

A rapid and easy to perform method for determining oxidized metallothionein (MT) is described. The main features of the procedure are that oxidized MT is converted into native MT with 2 -mercaptoethanol as reducing agent and (\mathrm{Zn}^{2+}) as metal donor, and MT is subsequently quantifled via (\mathrm{Cd}) saturation. The procedure was effective in recovering MT oxidized either by (\mathrm{Cu}^{2+}) or by neutralization of apothionein at (\mathrm{pH} 3), independent of the amount of MT, the origin of sample, degree of oxidation, and oxidation method. As demonstrated by HPLC analysis, the method is highly specific for MT. The described procedure provides information on both the total concentration of MT independent of its thiol redox state and the metal binding capacity of the protein; thus, it is of particular interest in studying the role of MT in metal metabolism and toxicity. (\sigma_{c} 1994) Academic Press, Inc.