Biochemical and immunological demonstration of prostaglandin D2, E2, and F2α formation from prostaglandin H2 by various rat glutathione S-transferase isozymes

doi:10.1016/0003-9861(88)90308-6

Archives of Biochemistry and Biophysics

Volume 264, Issue 2, August 1988, Pages 428-437

https://doi.org/10.1016/0003-9861(88)90308-6 Get rights and content

Abstract

Glutathione S-transferase isozymes purified from normal rat liver (1-1, 1–2, 2-2, 3-3, 3–4, and 4-4), liver with hyperplastic nodules (7-7), brain (Yn1Yn1), and testis (Yn1Yn2) all had prostaglandin H₂-converting activity. The prostaglandin H₂ E-isomerase activity was high in 1-1 (1400 nmol/min/mg protein), 1–2 (1170), and 2-2 (420), moderate in 3-3, 3–4, 4-4, Yn1Yn1, and Yn1Yn2 (52–100), and weak but significant in 7-7 (33). The prostaglandin H₂ D-isomerase activity was relatively high in 1-1 (170) and 1–2 (200), moderate in 2-2 (60) and Yn1Yn2 (43), and weak but marked in 3-3 (16), 4-4 (16), and 7-7 (14). The prostaglandin H₂ F-reductase activity was remarkable in 1-1 (1250), 1–2 (920), and 2-2 (390), and weakly detected in 3-3 (24), 4-4 (28), and 7-7 (14). Glutathione was absolutely required for these prostaglandin H₂-converting reactions, and its stoichiometric consumption was associated with F-reductase activity but not E- and D-isomerase activities. The K_m values for glutathione and prostaglandin H₂ were about 200 and 10–40 μm, respectively. By immunoabsorption analyses with various antibodies specific for each isozyme, we examined its contribution to the formation of prostaglandins D₂, E₂, and F_2α from prostaglandin H₂ in 100,000g supernatants of rat liver, kidney, and testis. In the liver, about 90% of the F-reductase activity (9.8 nmol/min/mg protein) was shown to be catalyzed by the 1–2 group of isozymes. The E-isomerase activity (16.5) was catalyzed about 60 and 40% by the 1–2 and 3–4 groups, respectively; and the D-isomerase activity (3.7) was catalyzed by the 1–2 group (50%) and the 3–4 group and Yn1Yn2 (15–25%). In the kidney, the E-isomerase activity (9.4) was catalyzed by 1-1, 1–2 (40%), 2-2, 3–4 group, and 7-7 (10–20%). The F-reductase activity (3.3) was mostly catalyzed by the 1–2 group (75%). In the testis, the E-isomerase activity (3.9) was catalyzed by the 1–2 group (20–30%), the 3–4 group, and Yn1Yn2 (30–60%).

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