Chemotherapy and Metabolic InhibitorsInteractions of prostaglandin A2 with the glutathione-mediated biotransformation system
Section snippets
Materials
GSH and NADH were obtained from Boehringer, Mannheim. Prostaglandins A2 and J2 were purchased from Sigma. CDNB was obtained from Aldrich Chemie. HPLC-grade trifluoroacetic acid was obtained from Baker. HPLC-grade methanol was from Labscan. DNPSG was synthesised analogously to Sokolovsky et al. [26].
[3H]Glycine was purchased from Amersham. GSTP1-1 was purified as previously described [27]. The three mutants of GSTP1-1, C47S, C101S, and C47S/C101S, were a generous gift from Dr. M. LoBello (Dept.
Inhibition studies with purified GSTP1-1
Time-dependent inhibition of GSTP1-1 and the mutants C47S, C101S, and C47S/C101S by prostaglandin A2 was studied. In Fig. 1A, the percentage of remaining GST activity after incubating 10, 25, or 250 μM of the prostaglandin A2 with GSTP1-1 is presented. Remaining activity after 3 hr was 74%, 56%, and 34%, respectively. Incubating these concentrations with the C47S mutant still resulted in a decrease in GST activity, albeit less evident than with the parent enzyme and only at the highest
Discussion
Prostaglandin A2 is known to inhibit cell proliferation 1, 2, 3, 4, 5, and numerous studies have been done on the metabolic fate of this prostaglandin. Recent findings concerning the formation and transport of the diastereoisomeric glutathione conjugates shed further light on this metabolism 20, 25. The present study aimed to obtain a more complete picture of the glutathione-related metabolism of PGA2 by investigating interactions with purified enzymes as well as in a cellular system using the
Acknowledgements
We thank Dr. M. LoBello for providing the GSTP1-1 mutants C47S, C101S, and C47S/C101S.
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2011, Journal of ProteomicsCitation Excerpt :In addition, cyPG adducts with immobilized thiols have been shown to be stable, being dissociated only by alkali treatment [10]. However, certain cyPG-protein adducts have been proposed to undergo retro-Michael reaction, like in the case of the GSTP1-1-PGA2 adduct [16]. Moreover, cyPG could interact in a reversible manner with some proteins.
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