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J Clin Invest. 1988 March; 81(3): 813–817.
PMCID: PMC442530

Defect in biosynthesis of mitochondrial acetoacetyl-coenzyme A thiolase in cultured fibroblasts from a boy with 3-ketothiolase deficiency.


The etiology of 3-ketothiolase deficiency has been attributed to a defect of mitochondrial acetoacetyl-CoA thiolase because the acetoacetyl-CoA thiolase activity in related materials is not activated by K+, a property characteristic for this enzyme. We studied the enzyme protein and the biosynthesis of mitochondrial acetoacetyl-CoA thiolase, using cultured skin fibroblasts from a 5-yr-old boy with 3-ketothiolase deficiency. The following results were obtained. (a) Activation of acetoacetyl-CoA thiolase activity by K+ was nil; (b) The enzyme activity was not affected by treatment with the antibody against mitochondrial acetoacetyl-CoA thiolase; (c) A signal for mitochondrial acetoacetyl-CoA thiolase protein was not detected in the immunoblot analysis; and (d) Pulse-chase experiments of skin fibroblasts, using [35S]methionine, revealed no incorporation of radioactivity into this enzyme. Therefore, fibroblasts from this patient lacked mitochondrial acetoacetyl-CoA thiolase protein due to a defect in its biosynthesis.

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Selected References

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  • Daum RS, Lamm PH, Mamer OA, Scriver CR. A "new" disorder of isoleucine catabolism. Lancet. 1971 Dec 11;2(7737):1289–1290. [PubMed]
  • Daum RS, Scriver CR, Mamer OA, Delvin E, Lamm P, Goldman H. An inherited disorder of isoleucine catabolism causing accumulation of alpha-methylacetoacetate and alpha-methyl-beta -hydroxybutyrate, and intermittent metabolic acidosis. Pediatr Res. 1973 Mar;7(3):149–160. [PubMed]
  • Gompertz D, Saudubray JM, Charpentier C, Bartlett K, Goodey PA, Draffan GH. A defect in l-isoleucine metabolism associated with alpha-methyl-beta-hydroxybutyric and alpha-methylacetoacetic aciduria: quantitative in vivo and in vitro studies. Clin Chim Acta. 1974 Dec 17;57(3):269–281. [PubMed]
  • Hillman RE, Keating JP. Beta-ketothiolase deficiency as a cause of the "ketotic hyperglycinemia syndrome". Pediatrics. 1974 Feb;53(2):221–225. [PubMed]
  • Halvorsen S, Stokke O, Jellum E. A variant form of 2-methyl-3-hydroxybutyric and 2-methylacetoacetic aciduria. Acta Paediatr Scand. 1979 Jan;68(1):123–128. [PubMed]
  • Robinson BH, Sherwood WG, Taylor J, Balfe JW, Mamer OA. Acetoacetyl CoA thiolase deficiency: a cause of severe ketoacidosis in infancy simulating salicylism. J Pediatr. 1979 Aug;95(2):228–233. [PubMed]
  • Schutgens RB, Middleton B, vd Blij JF, Oorthuys JW, Veder HA, Vulsma T, Tegelaers WH. Beta-ketothiolase deficiency in a family confirmed by in vitro enzymatic assays in fibroblasts. Eur J Pediatr. 1982 Sep;139(1):39–42. [PubMed]
  • Middleton B, Bartlett K. The synthesis and characterisation of 2-methylacetoacetyl coenzyme A and its use in the identification of the site of the defect in 2-methylacetoacetic and 2-methyl-3-hydroxybutyric aciduria. Clin Chim Acta. 1983 Mar 14;128(2-3):291–305. [PubMed]
  • Bennett MJ, Littlewood JM, MacDonald A, Pollitt RJ, Thompson J. A case of beta-ketothiolase deficiency. J Inherit Metab Dis. 1983;6(4):157–157. [PubMed]
  • Middleton B, Gray RG, Bennett MJ. Two cases of beta-ketothiolase deficiency: a comparison. J Inherit Metab Dis. 1984;7 (Suppl 2):131–132. [PubMed]
  • Middleton B, Bartlett K, Romanos A, Gomez Vazquez J, Conde C, Cannon RA, Lipson M, Sweetman L, Nyhan WL. 3-Ketothiolase deficiency. Eur J Pediatr. 1986 Apr;144(6):586–589. [PubMed]
  • Hiyama K, Sakura N, Matsumoto T, Kuhara T. Deficient beta-ketothiolase activity in leukocytes from a patient with 2-methylacetoacetic aciduria. Clin Chim Acta. 1986 Mar 16;155(2):189–194. [PubMed]
  • Middleton B. The oxoacyl-coenzyme A thiolases of animal tissues. Biochem J. 1973 Apr;132(4):717–730. [PubMed]
  • Miyazawa S, Osumi T, Hashimoto T. The presence of a new 3-oxoacyl-CoA thiolase in rat liver peroxisomes. Eur J Biochem. 1980 Feb;103(3):589–596. [PubMed]
  • Middleton B. The kinetic mechanism and properties of the cytoplasmic acetoacetyl-coenzyme A thiolase from rat liver. Biochem J. 1974 Apr;139(1):109–121. [PubMed]
  • Markwell MA, Haas SM, Bieber LL, Tolbert NE. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem. 1978 Jun 15;87(1):206–210. [PubMed]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. [PubMed]
  • Takiguchi M, Mori M, Tatibana M. A simple and rapid procedure for high-yield isolation of essentially undegraded free and membrane-bound polysomes from rat liver. J Biochem. 1985 May;97(5):1447–1459. [PubMed]
  • Tager JM, Van der Beek WA, Wanders RJ, Hashimoto T, Heymans HS, Van den Bosch H, Schutgens RB, Schram AW. Peroxisomal beta-oxidation enzyme proteins in the Zellweger syndrome. Biochem Biophys Res Commun. 1985 Feb 15;126(3):1269–1275. [PubMed]
  • Ozasa H, Furuta S, Miyazawa S, Osumi T, Hashimoto T, Mori M, Miura S, Tatibana M. Biosynthesis of enzymes of rat-liver mitochondrial beta-oxidation. Eur J Biochem. 1984 Nov 2;144(3):453–458. [PubMed]
  • Miura S, Mori M, Takiguchi M, Tatibana M, Furuta S, Miyazawa S, Hashimoto T. Biosynthesis and intracellular transport of enzymes of peroxisomal beta-oxidation. J Biol Chem. 1984 May 25;259(10):6397–6402. [PubMed]
  • Schram AW, Strijland A, Hashimoto T, Wanders RJ, Schutgens RB, van den Bosch H, Tager JM. Biosynthesis and maturation of peroxisomal beta-oxidation enzymes in fibroblasts in relation to the Zellweger syndrome and infantile Refsum disease. Proc Natl Acad Sci U S A. 1986 Aug;83(16):6156–6158. [PubMed]

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