D. Anuradha et al., Purification and characterization of rat testicular glutathione S-transferases: role in the synthesis of eicosanoids, ASIAN J AND, 2(4), 2000, pp. 277-282
Aim: Purification of glutathione S-transferases (GSTs) from rat testis; sep
aration and-identification of various subunits and their role in eicosanoid
biosynthesis. Methods: Purification of mt testicular GSTs by affinity chro
matography, employing S-hexylglutathione-linked epoxy-activated Sepharose 6
B column and separation of individual subunits by reverse phase-high pressu
re liquid chromatography (RP-HPLC). Characterization of affinity purified G
STs by Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)
and Western blot analysis. The role of testicular GSTs in eicosanoid biosy
nthesis was determined by incubating GSTs with 5,6-Leukotriene A(4)Me (LTA(
4)Me) and prostaglandin H-2(PGH(2)) and analyzing the products formed on HP
LC/TLC. Results: The present study reveals that majority of rat testicular
GSTs are of Y-b size (60%) with molecular weight of 27 kDa. The most predom
inant subunits, however, are GST Y-n2(27%), followed by GST Y-c(24%) and GS
T Y-n1(20%). These testicular GSTs showed very high Leukotriene C-4 (LTC4)
synthase activity with 5, 6-Leukotriene A(4)Me (LTA(4)Me) as the substrate
and prostaglandin D (PGD) synthase activity with prostaglandin H-2(PGH(2))
as the substrate. Conclusion: Majority of rat testicular GSTs are Y-b sized
and are involved in the synthesis of eicosanoids like LTC4 and PG(4).