Determination of glutathione-S-transferase traces in preparations of p53 C-terminal domain (aa320-393)

Bioelectrochemistry. 2002 Jan;55(1-2):115-8. doi: 10.1016/s1567-5394(01)00137-2.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Glutathione Transferase / analysis*
  • Potentiometry
  • Recombinant Fusion Proteins / chemistry*
  • Tumor Suppressor Protein p53 / chemistry*

Substances

  • Recombinant Fusion Proteins
  • Tumor Suppressor Protein p53
  • Glutathione Transferase