Expression of glutathione transferase isoenzymes in the porcine ovary in relationship to follicular maturation and luteinization.
The expression of different isoenzymes of glutathione transferase (GST), i.e. the cytosolic subunits GSTA1/A2, A3, A4, A5, M1/2, M2 and P1, T2, and the microsomal GST in follicles of different sizes and in corpora lutea from porcine ovary, was investigated by Western blotting. No immunoreactivity was obtained with anti-rat GSTT2 or anti-rat microsomal GST polyclonal antibodies. In contrast, GSTA1/A2, A3, A4, A5, M1/2, M2 and P1 are all expressed in the cytosol from porcine ovaries. In general, the highest levels of these GST isoenzymes were present in the cytosol from corpora lutea, in agreement with measurements of activity towards 1-chloro-2,4-dinitrobenzene. Immunoreactivity with anti-rat GSTP1 was only obtained with follicles. The cytosolic GSTs from follicles and corpora lutea were affinity purified on glutathione-Sepharose and separated by reversed-phase high-performance liquid chromatography in order to quantitate the different subunits. A peak corresponding to the class pi subunit was present in follicles. This peak was also seen with corpora lutea, although at very low level. There were four peaks containing class mu subunits. The remaining peaks were concluded to contain the class alpha subunits, except for two peaks which are suggested to contain proteins other than GSTs. The levels of the different subunits were quantitated on the basis of the areas under the peaks and the relative amounts in follicles of different sizes and in corpora lutea corresponded well with the Western blot analysis.
Eliasson
M
M
Department of Biochemistry, Wallenberg Laboratory, Stockholm University, Sweden. Maria@tuborg.biokemi.su.se
Stark
T
T
DePierre
J W
JW
D000818:Animals; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid / Q000662:veterinary; D003600:Cytosol / Q000201:enzymology; D005260:Female; D005982:Glutathione Transferase / Q000378:metabolism*; D007527:Isoenzymes / Q000378:metabolism; D008184:Luteal Cells / Q000201:enzymology*; D007986:Luteinizing Hormone / Q000378:metabolism*; D008861:Microsomes / Q000201:enzymology; D006080:Ovarian Follicle / Q000201:enzymology*; D013552:Swine / Q000502:physiology*
D007527:Isoenzymes; D007986:Luteinizing Hormone; D005982:Glutathione Transferase