Troglitazone stimulates acetyl-CoA carboxylase activity through a post-translational mechanism.
Troglitazone, a thiazolidinedione, is known to act as an insulin sensitizer. The various effects of the drug include stimulation of glucose utilization and inhibition of gluconeogenesis and fatty acid oxidation. We studied the effect of troglitazone treatment on rat liver acetyl-CoA carboxylase (ACC), the key enzyme that catalyzes the formation of malonyl-CoA, the rate-limiting step in the synthesis of long chain fatty acids. Treatment of rats with troglitazone for 18 days resulted in more than 200% increase in the activity of hepatic acetyl-CoA carboxylase (1.01+/-0.14 and 2.33+/-0.28 mU/mg supernatant protein for control and troglitazone-treated rats, respectively) (p<0.001). The expression of acetyl-CoA carboxylase mRNA, as studied by RNAse protection assay, was not significantly different between the two groups of animals. The ACC from control and troglitazone-treated groups was purified by avidin-affinity chromatography. The purified enzyme migrated as a major protein band (Mr 262,000) on SDS-polyacrylamide gels. Troglitazone treatment was associated with increased citrate sensitivity of ACC. The specific activity of the purified preparation in troglitazone-treated rats was increased by 67% (2.5 vs. 1.5 U/mg). Quantitation of alkali-labile phosphate content of the purified preparation revealed 5.66+/-0.17 and 6.29+/-0.13 mol Pi/mol subunit of 262 Kda for control and troglitazone-treated rats, respectively (P<0.01). The subtle increase in phosphate content does not explain the observed activation of the enzyme. It is possible that additional mechanisms such as troglitazone related rearrangement of the occupancy of select phosphate binding sites or altered binding of the biotin cofactor may also contribute to the observed activation of ACC.
Thampy
G K
GK
Department of Internal Medicine, Saint Louis University School of Medicine, Missouri 63110, USA.
Haas
M J
MJ
Mooradian
A D
AD
D000103:Acetyl-CoA Carboxylase / Q000096:biosynthesis*; D000818:Animals; D001835:Body Weight / Q000187:drug effects; D002839:Chromans / Q000494:pharmacology*; D004435:Eating / Q000187:drug effects; D007004:Hypoglycemic Agents / Q000494:pharmacology*; D007202:Indicators and Reagents; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008297:Male; D023282:Nuclease Protection Assays; D010710:Phosphates / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D011916:Rats, Inbred F344; D013268:Stimulation, Chemical; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D013844:Thiazoles / Q000494:pharmacology*; D045162:Thiazolidinediones; D000077288:Troglitazone
D002839:Chromans; D007004:Hypoglycemic Agents; D007202:Indicators and Reagents; D010710:Phosphates; D012333:RNA, Messenger; D013844:Thiazoles; D045162:Thiazolidinediones; D000103:Acetyl-CoA Carboxylase; D000077288:Troglitazone