Effect of glutathione deficiency on the adipocyte sn-glycerol-3-phosphate acyltransferase
| Autor: | Subhash C. Jamdar, Edward Soo, Wang Fang Cao |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Male
Biophysics Maleates Adipose tissue Glutathione Biochemistry Dithiothreitol Mitochondria Rats Rats Sprague-Dawley chemistry.chemical_compound Endocrinology chemistry In vivo Adipocyte Acyltransferase Microsomes Glycerol-3-Phosphate O-Acyltransferase Microsome Adipocytes Animals Buthionine sulfoximine Buthionine Sulfoximine |
| Zdroj: | Biochimica et biophysica acta. 1393(1) |
| ISSN: | 0006-3002 |
| Popis: | The present study investigates the effects of various glutathione (GSH) depleting agents on sn-glycerol-3-phosphate acyltransferase (GPAT) activity, the first committed step in adipose triacylglycerol formation. GPAT activity was measured in the presence of [14C]glycerol-3-phosphate and palmitoyl-CoA, using different subcellular fractions. Glutathione deficiency in animals was induced in the presence of diethylmaleate (DEM) or buthionine sulfoximine. In this respect, DEM (1.75 mmoles/kg) was more effective and caused over 75% decrease in GPAT activity within 4 h of DEM administration. Further studies indicated that this decrease in GPAT activity was mainly related to the microsomal form of GPAT, without any significant effect on mitochondrial GPAT activity. Adipocytes incubated with 2.5 mm DEM for 1 h at 37 degrees C also showed a reduction in the adipocyte glutathione content, which was accompanied by decreases in GPAT activity. The effect of DEM on adipocyte GPAT activity was partially reversible in the presence of cell permeable glutathione ethyl ester. Preincubation of adipose tissue homogenates with 2.5 mM DEM at 30 degrees C for 45 min also showed a significant loss of the GPAT activity. The presence of 5 mM dithiothreitol in the preincubation mixture offered a significant protection of the GPAT activity against DEM. However, glutathione was ineffective in this respect as it interfered with the utilization of palmitoyl-CoA in the GPAT assay. Therefore, on the basis of these three different approaches, the present studies suggest that the thiol environment offered by glutathione (in vivo and in vitro studies) or dithiothreitol (in a cell-free system) is critical for the maintenance of GPAT activity. |
| Databáze: | OpenAIRE |
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