Determination of glutathione-S-transferase traces in preparations of p53 C-terminal domain (aa320–393)
✍ Scribed by Marie Brázdová; René Kizek; Ludek Havran; Emil Paleček
- Book ID
- 104443885
- Publisher
- Elsevier Science
- Year
- 2002
- Tongue
- English
- Weight
- 83 KB
- Volume
- 55
- Category
- Article
- ISSN
- 1567-5394
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✦ Synopsis
Tumor suppressor protein p53 is often expressed as a fusion protein with glutathione-S-transferase (GST). The sensitive determination of GST in p53 samples is thus necessary. We propose a method for the determination of traces of GST in the p53 C-terminus based on the constant current chronopotentiometric stripping analysis (CPSA) with hanging mercury drop electrode (HMDE). GST produces a catalytic signal in cobalt-containing solutions due to cysteine residues. A large excess of the C-terminus does interfere with the determination because of the lack of cysteines in the molecule. This method is simple and very sensitive and is capable of detecting < 1% GST in the p53 sample.