title: Counter modulation of adipocyte mitochondrial processes by insulin and S-oxalylglutathione.
abstract: Oxalyl thiolesters, a group of putative intracellular regulators, have been shown to be in vitro inhibitors of some cytosolic enzymes which are stimulated by insulin. In this study, the effects of insulin and oxalyl thiolesters on pyruvate dehydrogenase, beta-oxidation, and acyl-CoA hydrolase activities in mitochondria from rat epididymal adipocytes are compared. Using glutathione, CoASH, cysteine, and cysteamine as thiol sources, oxalyl thioesters were synthesized, purified, and quantitated. Mitochondria were isolated from rat epididymal adipocytes, some of which were incubated with or without insulin. Mitochondrial activities were determined by radioisotopic assay subsequent to control, insulin, or oxalyl thiolester incubation. Under the conditions used in this study, pyruvate dehydrogenase activity was increased 28% subsequent to 10-min incubation of adipocytes with 400 microU/ml insulin; in contrast, preincubation of adipocyte mitochondria with S-oxalylglutathione resulted in a dose-dependent 11-19% inhibition of pyruvate dehydrogenase. S-oxalylglutathione also attenuated the spermine-induced activation of pyruvate dehydrogenase. Insulin treatment resulted in a small but significant increase in beta-oxidation of palmitic acid while 100 microM S-oxalylglutathione mediated a 40% decrease in palmitate oxidation. Palmitoyl-CoA hydrolase activity was decreased 14% by insulin treatment; however, S-oxalylglutathione caused a 14-50% increase in hydrolase activity. The other oxalyl thiolesters were not as effective or as consistent as S-oxalylglutathione in modulation of the mitochondrial activities; free thiols and oxalic acid did not modulate the activities. In summary, pyruvate dehydrogenase, palmitate beta-oxidation, and palmitoyl-CoA hydrolase activities in adipocyte mitochondria were modulated in approximately equal but opposite directions by insulin and S-oxalylglutathione. These findings support the suggestion that oxalyl thiolesters may function as an intracellular signal recruited to return insulin to normal levels.
authors.affiliation: Department of Chemistry, Oakland University, Rochester, MI 48309-4401, USA.
authors.forename: K H
authors.initials: KH
authors.lastname: Moore
authors.forename: P
authors.initials: P
authors.lastname: Tsatsos
authors.forename: D M
authors.initials: DM
authors.lastname: Staudacher
authors.forename: F L
authors.initials: FL
authors.lastname: Kiechle
mesh_terms: D017667:Adipocytes / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D000818:Animals; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004822:Epididymis / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005978:Glutathione / Q000031:analogs & derivatives* / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology*; D008297:Male; D008928:Mitochondria / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D010084:Oxidation-Reduction; D010170:Palmitoyl-CoA Hydrolase / Q000187:drug effects; D011768:Pyruvate Dehydrogenase Complex / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D004791:Enzyme Inhibitors; D007328:Insulin; D011768:Pyruvate Dehydrogenase Complex; C052145:S-oxalylglutathione; D010170:Palmitoyl-CoA Hydrolase; D005978:Glutathione