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. 1997 Apr 15;323(Pt 2):525–531. doi: 10.1042/bj3230525

Peroxisome proliferator-induced acyl-CoA thioesterase from rat liver cytosol: molecular cloning and functional expression in Chinese hamster ovary cells.

S T Engberg 1, T Aoyama 1, S E Alexson 1, T Hashimoto 1, L T Svensson 1
PMCID: PMC1218351  PMID: 9163348

Abstract

We have isolated and cloned a cDNA that codes for one of the peroxisome proliferator-induced acyl-CoA thioesterases of rat liver. The deduced amino acid sequence corresponds to the major induced isoform in cytosol. Analysis and comparison of the deduced amino acid sequence with the established consensus sequences suggested that this enzyme represents a novel kind of esterase with an incomplete lipase serine active site motif. Analyses of mRNA and its expression indicated that the enzyme is significantly expressed in liver only after peroxisome proliferator treatment, but isoenzymes are constitutively expressed at high levels in testis and brain. The reported cDNA sequence is highly homologous to the recently cloned brain acyl-CoA thioesterase [Broustas, Larkins, Uhler and Hajra (1996) J. Biol. Chem. 271, 10470-10476], but subtle differences throughout the sequence, and distinct differences close to the resulting C-termini, suggest that they are different enzymes, regulated in different manners. A full-length cDNA clone was expressed in Chinese hamster ovary cells and the expressed enzyme was characterized. The palmitoyl-CoA hydrolysing activity (Vmax) was induced approx. 9-fold to 1 micromol/min per mg of cell protein, which was estimated to correspond to a specific activity of 250 micromol/min per mg of cDNA-expressed enzyme. Both the specific activity and the acyl-CoA chain length specificity were very similar to those of the purified rat liver enzyme.

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Selected References

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  1. Alexson S. E., Mentlein R., Wernstedt C., Hellman U. Isolation and characterization of microsomal acyl-CoA thioesterase. A member of the rat liver microsomal carboxylesterase multi-gene family. Eur J Biochem. 1993 Jun 15;214(3):719–727. doi: 10.1111/j.1432-1033.1993.tb17973.x. [DOI] [PubMed] [Google Scholar]
  2. Aoyama T., Ueno I., Kamijo T., Hashimoto T. Rat very-long-chain acyl-CoA dehydrogenase, a novel mitochondrial acyl-CoA dehydrogenase gene product, is a rate-limiting enzyme in long-chain fatty acid beta-oxidation system. cDNA and deduced amino acid sequence and distinct specificities of the cDNA-expressed protein. J Biol Chem. 1994 Jul 22;269(29):19088–19094. [PubMed] [Google Scholar]
  3. BARTLETT G. R. Phosphorus assay in column chromatography. J Biol Chem. 1959 Mar;234(3):466–468. [PubMed] [Google Scholar]
  4. Broustas C. G., Hajra A. K. Purification, properties, and specificity of rat brain cytosolic fatty acyl coenzyme A hydrolase. J Neurochem. 1995 May;64(5):2345–2353. doi: 10.1046/j.1471-4159.1995.64052345.x. [DOI] [PubMed] [Google Scholar]
  5. Broustas C. G., Larkins L. K., Uhler M. D., Hajra A. K. Molecular cloning and expression of cDNA encoding rat brain cytosolic acyl-coenzyme A thioester hydrolase. J Biol Chem. 1996 May 3;271(18):10470–10476. doi: 10.1074/jbc.271.18.10470. [DOI] [PubMed] [Google Scholar]
  6. Camp L. A., Hofmann S. L. Purification and properties of a palmitoyl-protein thioesterase that cleaves palmitate from H-Ras. J Biol Chem. 1993 Oct 25;268(30):22566–22574. [PubMed] [Google Scholar]
  7. Camp L. A., Verkruyse L. A., Afendis S. J., Slaughter C. A., Hofmann S. L. Molecular cloning and expression of palmitoyl-protein thioesterase. J Biol Chem. 1994 Sep 16;269(37):23212–23219. [PubMed] [Google Scholar]
  8. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  9. Derewenda Z. S., Sharp A. M. News from the interface: the molecular structures of triacylglyceride lipases. Trends Biochem Sci. 1993 Jan;18(1):20–25. doi: 10.1016/0968-0004(93)90082-x. [DOI] [PubMed] [Google Scholar]
  10. Issemann I., Green S. Activation of a member of the steroid hormone receptor superfamily by peroxisome proliferators. Nature. 1990 Oct 18;347(6294):645–650. doi: 10.1038/347645a0. [DOI] [PubMed] [Google Scholar]
  11. Kamijo T., Aoyama T., Miyazaki J., Hashimoto T. Molecular cloning of the cDNAs for the subunits of rat mitochondrial fatty acid beta-oxidation multienzyme complex. Structural and functional relationships to other mitochondrial and peroxisomal beta-oxidation enzymes. J Biol Chem. 1993 Dec 15;268(35):26452–26460. [PubMed] [Google Scholar]
  12. Katoh H., Nakajima S., Kawashima Y., Kozuka H., Uchiyama M. Induction of rat hepatic long-chain acyl-CoA hydrolases by various peroxisome proliferators. Biochem Pharmacol. 1984 Apr 1;33(7):1081–1085. doi: 10.1016/0006-2952(84)90517-3. [DOI] [PubMed] [Google Scholar]
  13. Kawashima Y., Katoh H., Kozuka H. Differential effects of altered hormonal state on the induction of acyl-CoA hydrolases and peroxisomal beta-oxidation by clofibric acid. Biochim Biophys Acta. 1983 Feb 7;750(2):365–372. doi: 10.1016/0005-2760(83)90041-3. [DOI] [PubMed] [Google Scholar]
  14. Kawashima Y., Katoh H., Kozuka H. Sex-related difference in the effect of clofibric acid on induction of two novel long-chain acyl-CoA hydrolases in rat liver. Biochim Biophys Acta. 1982 Jul 20;712(1):48–56. [PubMed] [Google Scholar]
  15. Lehner R., Kuksis A. Purification of an acyl-CoA hydrolase from rat intestinal microsomes. A candidate acyl-enzyme intermediate in glycerolipid acylation. J Biol Chem. 1993 Nov 25;268(33):24726–24733. [PubMed] [Google Scholar]
  16. Mentlein R., Rix-Matzen H., Heymann E. Subcellular localization of non-specific carboxylesterases, acylcarnitine hydrolase, monoacylglycerol lipase and palmitoyl-CoA hydrolase in rat liver. Biochim Biophys Acta. 1988 Mar 17;964(3):319–328. doi: 10.1016/0304-4165(88)90032-3. [DOI] [PubMed] [Google Scholar]
  17. Miyazawa S., Furuta S., Hashimoto T. Induction of a novel long-chain acyl-CoA hydrolase in rat liver by administration of peroxisome proliferators. Eur J Biochem. 1981 Jul;117(2):425–430. doi: 10.1111/j.1432-1033.1981.tb06356.x. [DOI] [PubMed] [Google Scholar]
  18. Naggert J., Williams B., Cashman D. P., Smith S. Cloning and sequencing of the medium-chain S-acyl fatty acid synthetase thioester hydrolase cDNA from rat mammary gland. Biochem J. 1987 Apr 15;243(2):597–601. doi: 10.1042/bj2430597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Naggert J., Witkowski A., Mikkelsen J., Smith S. Molecular cloning and sequencing of a cDNA encoding the thioesterase domain of the rat fatty acid synthetase. J Biol Chem. 1988 Jan 25;263(3):1146–1150. [PubMed] [Google Scholar]
  20. Niwa H., Yamamura K., Miyazaki J. Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene. 1991 Dec 15;108(2):193–199. doi: 10.1016/0378-1119(91)90434-d. [DOI] [PubMed] [Google Scholar]
  21. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Smith S. The animal fatty acid synthase: one gene, one polypeptide, seven enzymes. FASEB J. 1994 Dec;8(15):1248–1259. [PubMed] [Google Scholar]
  23. Sugiyama E., Uemura K., Hara A., Taketomi T. Effects of various lysosphingolipids on cell growth, morphology and lipid composition in three neuroblastoma cell lines. Biochem Biophys Res Commun. 1990 Jun 15;169(2):673–679. doi: 10.1016/0006-291x(90)90383-x. [DOI] [PubMed] [Google Scholar]
  24. Svensson L. T., Alexson S. E., Hiltunen J. K. Very long chain and long chain acyl-CoA thioesterases in rat liver mitochondria. Identification, purification, characterization, and induction by peroxisome proliferators. J Biol Chem. 1995 May 19;270(20):12177–12183. doi: 10.1074/jbc.270.20.12177. [DOI] [PubMed] [Google Scholar]
  25. Svensson L. T., Wilcke M., Alexson S. E. Peroxisome proliferators differentially regulate long-chain acyl-CoA thioesterases in rat liver. Eur J Biochem. 1995 Jun 1;230(2):813–820. doi: 10.1111/j.1432-1033.1995.0813h.x. [DOI] [PubMed] [Google Scholar]
  26. Yamada J., Furihata T., Tamura H., Watanabe T., Suga T. Long-chain acyl-CoA hydrolase from rat brain cytosol: purification, characterization, and immunohistochemical localization. Arch Biochem Biophys. 1996 Feb 1;326(1):106–114. doi: 10.1006/abbi.1996.0053. [DOI] [PubMed] [Google Scholar]
  27. Yamada J., Matsumoto I., Furihata T., Sakuma M., Suga T. Purification and properties of long-chain acyl-CoA hydrolases from the liver cytosol of rats treated with peroxisome proliferator. Arch Biochem Biophys. 1994 Jan;308(1):118–125. doi: 10.1006/abbi.1994.1017. [DOI] [PubMed] [Google Scholar]

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