Quantitative profiling of tissue- and gender-related expression of glutathione S-transferase isoenzymes in the mouse.
Cytosolic glutathione S-transferase (GST) isoenzymes from brain, heart, lung, liver, kidney and gonads of male and female CD-1 mice were identified and quantified with a combination of affinity purification, electrospray ionization MS, Edman microsequencing, Western blot analysis and reverse-phase HPLC. The three principal hepatic GST subunits, mGSTA3 (25271 Da), mGSTP1 (23478 Da), and mGSTM1 (25839 Da), were isolated from liver, lung, kidney, testes and female heart, whereas brain, ovaries and male heart contained mGSTM1 and mGSTP1. Additional isoenzymes were detected in tissues, including mu class subunits mGSTM2 (25580 Da) and mGSTM3 (25570 Da), an N-terminally blocked Alpha subunit (25480 Da) assigned as mGSTA4, and proteins of molecular masses 25490, 22540, 24493, 24378 and 25383 Da. Distinct gender differences in expression of GST subunits were observed for liver, heart, kidney and gonads, whereas GST expression was similar in brain and lung for both genders. In contrast with patterns of expression in liver (high ratio of mGSTA3 to mGSTP1 in females relative to males), mGSTP1 was the most abundant subunit in female gonads, whereas mGSTA3 was not present in detectable quantities. The profile of GST expression in kidney was similar to that in liver; however, male kidneys expressed 30% more soluble GST than female kidneys. A striking gender-related difference in GST expression was found in cardiac tissue, where female animals expressed 50% more soluble GST than male tissues, and the GST isoenzyme with the greatest documented activity towards lipid hydroperoxides, mGSTA3, was present in female tissue yet absent from male tissue. These results point to complex gender- and tissue-dependent expression of individual mouse GST isoenzymes.
Mitchell
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AE
Facility for Advanced Instrumentation, University of California, Davis, CA 95616-8597, USA.
Morin
D
D
Lakritz
J
J
Jones
A D
AD
D000818:Animals; D001921:Brain / Q000201:enzymology; D002851:Chromatography, High Pressure Liquid; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D005982:Glutathione Transferase / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D007527:Isoenzymes / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D046911:Macromolecular Substances; D008297:Male; D051379:Mice; D009206:Myocardium / Q000201:enzymology; D009928:Organ Specificity; D010053:Ovary / Q000201:enzymology; D012727:Sex Characteristics; D018629:Spectrometry, Mass, Secondary Ion; D013737:Testis / Q000201:enzymology
D007527:Isoenzymes; D046911:Macromolecular Substances; D005982:Glutathione Transferase