Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1996 Aug 15;98(4):1028–1033. doi: 10.1172/JCI118863

Common missense mutation G1528C in long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency. Characterization and expression of the mutant protein, mutation analysis on genomic DNA and chromosomal localization of the mitochondrial trifunctional protein alpha subunit gene.

L IJlst 1, J P Ruiter 1, J M Hoovers 1, M E Jakobs 1, R J Wanders 1
PMCID: PMC507519  PMID: 8770876

Abstract

Mitochondrial trifunctional protein (MTP) is a recently identified enzyme involved in mitochondrial beta-oxidation, harboring long-chain enoyl-CoA hydratase, long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) and long-chain 3-ketothiolase activity. A deficiency of this protein is associated with impaired oxidation of long-chain fatty acids which can lead to sudden infant death. Furthermore, it is clear that this inborn error of fatty acid oxidation is very frequent, second to medium chain acyl-CoA dehydrogenase deficiency. In most patients only the LCHAD activity of MTP is deficient with near normal activity of the two other enzyme activities of the complex. We recently described the occurrence of a frequent G1528C mutation in the cDNA coding for the a subunit of MTP. Using S. cerevisiae for expression of wild type and mutant protein we show that the G1528C mutation is directly responsible for the loss of LCHAD activity. Furthermore, we describe a newly developed method allowing identification of the G1528C mutation in genomic DNA. The finding of an 87% allele frequency of the G1528C mutation in 34 LCHAD deficient patients makes this a valuable test for prenatal diagnosis. Finally, we show that the gene encoding the alpha subunit of MTP is located on chromosome 2p24.1-23.3.

Full Text

The Full Text of this article is available as a PDF (246.0 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Boom R., Sol C. J., Salimans M. M., Jansen C. L., Wertheim-van Dillen P. M., van der Noordaa J. Rapid and simple method for purification of nucleic acids. J Clin Microbiol. 1990 Mar;28(3):495–503. doi: 10.1128/jcm.28.3.495-503.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Carpenter K., Pollitt R. J., Middleton B. Human liver long-chain 3-hydroxyacyl-coenzyme A dehydrogenase is a multifunctional membrane-bound beta-oxidation enzyme of mitochondria. Biochem Biophys Res Commun. 1992 Mar 16;183(2):443–448. doi: 10.1016/0006-291x(92)90501-b. [DOI] [PubMed] [Google Scholar]
  3. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  4. Hiltunen J. K., Wenzel B., Beyer A., Erdmann R., Fosså A., Kunau W. H. Peroxisomal multifunctional beta-oxidation protein of Saccharomyces cerevisiae. Molecular analysis of the fox2 gene and gene product. J Biol Chem. 1992 Apr 5;267(10):6646–6653. [PubMed] [Google Scholar]
  5. IJlst L., Wanders R. J., Ushikubo S., Kamijo T., Hashimoto T. Molecular basis of long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency: identification of the major disease-causing mutation in the alpha-subunit of the mitochondrial trifunctional protein. Biochim Biophys Acta. 1994 Dec 8;1215(3):347–350. doi: 10.1016/0005-2760(94)90064-7. [DOI] [PubMed] [Google Scholar]
  6. Jackson S., Kler R. S., Bartlett K., Briggs H., Bindoff L. A., Pourfarzam M., Gardner-Medwin D., Turnbull D. M. Combined enzyme defect of mitochondrial fatty acid oxidation. J Clin Invest. 1992 Oct;90(4):1219–1225. doi: 10.1172/JCI115983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kamijo T., Aoyama T., Komiyama A., Hashimoto T. Structural analysis of cDNAs for subunits of human mitochondrial fatty acid beta-oxidation trifunctional protein. Biochem Biophys Res Commun. 1994 Mar 15;199(2):818–825. doi: 10.1006/bbrc.1994.1302. [DOI] [PubMed] [Google Scholar]
  8. Mu F. T., Baldwin G., Weinstock J., Stockman D., Toh B. H. Monoclonal antibody to the gastrin receptor on parietal cells recognizes a 78-kDa protein. Proc Natl Acad Sci U S A. 1987 May;84(9):2698–2702. doi: 10.1073/pnas.84.9.2698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Pollitt R. J. Disorders of mitochondrial long-chain fatty acid oxidation. J Inherit Metab Dis. 1995;18(4):473–490. doi: 10.1007/BF00710058. [DOI] [PubMed] [Google Scholar]
  10. Przyrembel H., Jakobs C., IJlst L., de Klerk J. B., Wanders R. J. Long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency. J Inherit Metab Dis. 1991;14(5):674–680. doi: 10.1007/BF01799932. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Sewell A. C., Bender S. W., Wirth S., Münterfering H., Ijlist L., Wanders R. J. Long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency: a severe fatty acid oxidation disorder. Eur J Pediatr. 1994 Oct;153(10):745–750. doi: 10.1007/BF01954492. [DOI] [PubMed] [Google Scholar]
  13. Sims H. F., Brackett J. C., Powell C. K., Treem W. R., Hale D. E., Bennett M. J., Gibson B., Shapiro S., Strauss A. W. The molecular basis of pediatric long chain 3-hydroxyacyl-CoA dehydrogenase deficiency associated with maternal acute fatty liver of pregnancy. Proc Natl Acad Sci U S A. 1995 Jan 31;92(3):841–845. doi: 10.1073/pnas.92.3.841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Treem W. R., Rinaldo P., Hale D. E., Stanley C. A., Millington D. S., Hyams J. S., Jackson S., Turnbull D. M. Acute fatty liver of pregnancy and long-chain 3-hydroxyacyl-coenzyme A dehydrogenase deficiency. Hepatology. 1994 Feb;19(2):339–345. [PubMed] [Google Scholar]
  15. Uchida Y., Izai K., Orii T., Hashimoto T. Novel fatty acid beta-oxidation enzymes in rat liver mitochondria. II. Purification and properties of enoyl-coenzyme A (CoA) hydratase/3-hydroxyacyl-CoA dehydrogenase/3-ketoacyl-CoA thiolase trifunctional protein. J Biol Chem. 1992 Jan 15;267(2):1034–1041. [PubMed] [Google Scholar]
  16. Wanders R. J., Duran M., Ijlst L., de Jager J. P., van Gennip A. H., Jakobs C., Dorland L., van Sprang F. J. Sudden infant death and long-chain 3-hydroxyacyl-CoA dehydrogenase. Lancet. 1989 Jul 1;2(8653):52–53. doi: 10.1016/s0140-6736(89)90300-0. [DOI] [PubMed] [Google Scholar]
  17. Wanders R. J., IJlst L., Poggi F., Bonnefont J. P., Munnich A., Brivet M., Rabier D., Saudubray J. M. Human trifunctional protein deficiency: a new disorder of mitochondrial fatty acid beta-oxidation. Biochem Biophys Res Commun. 1992 Nov 16;188(3):1139–1145. doi: 10.1016/0006-291x(92)91350-y. [DOI] [PubMed] [Google Scholar]
  18. Wilcken B., Leung K. C., Hammond J., Kamath R., Leonard J. V. Pregnancy and fetal long-chain 3-hydroxyacyl coenzyme A dehydrogenase deficiency. Lancet. 1993 Feb 13;341(8842):407–408. doi: 10.1016/0140-6736(93)92993-4. [DOI] [PubMed] [Google Scholar]
  19. Yaffe M. P. Analysis of mitochondrial function and assembly. Methods Enzymol. 1991;194:627–643. doi: 10.1016/0076-6879(91)94046-f. [DOI] [PubMed] [Google Scholar]
  20. Zhang Q. X., Baldwin G. S. Structures of the human cDNA and gene encoding the 78 kDa gastrin-binding protein and of a related pseudogene. Biochim Biophys Acta. 1994 Oct 18;1219(2):567–575. doi: 10.1016/0167-4781(94)90091-4. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES