Abstract
The tight binding of Meisenheimer intermediate with octopus digestive gland glutathione transferase was analyzed with 1,3,5-trinitrobenzene, which forms a trapped Meisenheimer complex with glutathione because there is no leaving group at the ipso carbon. By steady-state enzyme kinetic analysis, an inhibition constant of 1.89 ± 0.17 μM was found for the transient formed, S-(2,4,6-trinitrophenyl) glutathione. The above inhibition constant is 407-fold smaller than the K m value for the substrate (2,4-dinitrochlorobenzene). Thus, S-(2,4,6-trinitrophenyl) glutathione is considered to be a transition-state analog. The tight binding of this inhibitor to the enzyme provides an explanation for the involvement of the biological binding effect on the rate enhancement in the glutathione transferase-catalyzed SNAr mechanism.
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Liou, JY., Huang, TM. & Chang, GG. Inhibition of Octopus Glutathione Transferase by Meisenheimer Complex Analog, S-(2,4,6-trinitrophenyl) Glutathione. J Protein Chem 19, 615–620 (2000). https://doi.org/10.1023/A:1007195130725
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DOI: https://doi.org/10.1023/A:1007195130725