Determination of glutathione-S-transferase traces in preparations of p53 C-terminal domain (aa320-393).
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.
Brázdová
Marie
M
Institute of Biophysics, Academy of Sciences of the Czech Republic, Královopolská 135, CZ-612 65 Brno, Czech Republic.
Kizek
René
R
Havran
Ludek
L
Palecek
Emil
E
D005982:Glutathione Transferase / Q000032:analysis*; D011199:Potentiometry; D011993:Recombinant Fusion Proteins / Q000737:chemistry*; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry*
D011993:Recombinant Fusion Proteins; D016159:Tumor Suppressor Protein p53; D005982:Glutathione Transferase