Abstract
Trifunctional protein (TFP) plays a significant role in the mitochondrial β-oxidation of long-chain fatty acids. Its deficiency impairs the energy generating function of this pathway and causes hypoketotic hypoglycemia once hepatic glycogen stores are depleted. A Reye-like syndrome, cardiomyopathy, and sudden death may follow. The diagnosis is based on demonstration of significantly decreased enzyme activity of at least two of the three involved enzymes in fibroblasts. The possibility of prospective diagnosis of TFP deficiency by newborn screening using tandem mass spectrometry (MS/MS) has not been evaluated. We report the postmortem diagnosis of a male newborn, who suffered sudden death at 2 wk of age, and his younger sister, who died of cardiomyopathy complicated by acute heart failure at the age of 6 mo, after she had acquired a common viral infection. Blood spots from the original newborn screening cards were the only remaining material from the patients. Analysis by MS/MS revealed acylcarnitine profiles consistent with either TFP or long-chain 3-hydroxyacyl coenzyme A dehydrogenase (LCHAD) deficiency. To prove the diagnosis, the α- and β-subunit genes coding for TFP were examined. The patients were compound heterozygous for a 4-bp-deletion and an a→g missense mutation, both in the same exon 3 donor consensus splice site. This is the first report of the diagnosis of TFP deficiency using blood spots obtained for newborn screening and suggests that TFP deficiency may be detectable by prospective newborn screening using MS/MS.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- TFP:
-
mitochondrial trifunctional protein deficiency
- MS/MS:
-
tandem mass spectrometry
- LCHAD:
-
long-chain 3-hydroxyacyl coenzyme A dehydrogenase
References
Grünewald S, Bakkeren J, Wanders RA, Wendel U 1997 Neonatal lethal mitochondrial trifunctional protein deficiency mimicking a respiratory chain defect. J Inherit Metab Dis 20: 835–836
Roe CR, Coates PM 1995 Mitochondrial fatty acid oxidation disorders. In: Scriver CR, Beaudet AL, Sly WS, Valle D (eds) The metabolic and molecular bases of inherited disease. McGraw-Hill, New York, pp 1501–1533
Miyajima H, Orii KE, Shindo Y, Hashimoto T, Shinka T, Kuhara T, Matsumoto I, Shimizu H, Kaneko E 1997 Mitochondrial trifunctional protein deficiency associated with recurrent myoglobinuria in adolescence. Neurology 49: 833–837
Schaefer J, Jackson S, Dick DJ, Turnbull DM 1996 Trifunctional enzyme deficiency: adult presentation of a usually fatal β-oxidation defect. Ann Neurol 40: 597–602
Cederbaum SD 1998 SIDS and disorders of fatty acid oxidation: where do we go from here?. J Pediatr 132: 913–914
Sweetman L 1996 Newborn screening by tandem mass spectrometry. Clin Chem 42: 345–346
Rashed MS, Bucknall MP, Little D, Awad A, Jacob M, Alamoudi M, Alwattar M, Ozand PT 1997 Screening blood spots for inborn errors of metabolism by electrospray tandem mass spectrometry with a microplate batch process and a computer algorithm for automated flagging of abnormal profiles. Clin Chem 43: 1129–1141
Ziadeh R, Hoffman EP, Finegold DM, Hoop RC, Brackett JC, Strauss AW, Naylor EW 1995 Medium chain acyl-CoA dehydrogenase deficiency in Pennsylvania: neonatal screening shows high incidence and unexpected mutations frequencies. Pediatr Res 37: 675–678
Chace DH, Hillman SL, Millington DS, Kahler SG, Adam BW, Levy HL 1996 Rapid diagnosis of homocystinuria and other hypermethioninemias from newborns' blood spots by tandem mass spectrometry. Clin Chem 42: 349–355
Chace DH, Hillman SL, Millington DS, Kahler SG, Roe CR, Naylor EW 1995 Rapid diagnosis of maple syrup urine disease in blood spots from newborns by tandem mass spectrometry. Clin Chem 41: 62–68
Chace DH, Hillman SL, Van Hove JL, Naylor EW 1997 Rapid diagnosis of MCAD deficiency: quantitative analysis of octanoylcarnitine and other acylcarnitines in newborn blood spots by tandem mass spectrometry. Clin Chem 43: 2106–2113
Chace DH, Millington DS, Terada N, Kahler SG, Roe CR, Hofman LF 1993 Rapid diagnosis of phenylketonuria by quantitative analysis for phenylalanine and tyrosine in neonatal blood spots by tandem mass spectrometry. Clin Chem 39: 66–71
Kamijo T, Aoyama T, Komiyama A, Hashimoto T 1993 Molecular cloning of the cDNAs for the subunits of rat mitochondrial fatty acid beta-oxidation multienzyme complex. J Biol Chem 268: 26452–26460
Brackett JC, Sims HF, Rinaldo P, Shapiro S, Powell CK, Bennett MJ, Strauss AW 1995 Two α subunit donor splice site mutations cause human trifunctional protein deficiency. J Clin Invest 95: 2076–2082
Orii KE, Aoyama T, Wakui K, Fukushima Y, Miyajima H, Yamaguchi S, Orii T, Kondo N, Hashimoto T 1997 Genomic and mutational analysis of the mitochondrial trifunctional protein beta-subunit (HADHB) gene in patients with trifunctional protein deficiency. Hum Mol Genet 6: 1215–1224
Sims HF, Brackett JC, Powell CK, Treem WR, Hale DE, Bennett MJ, Gibson B, Shapiro S, Strauss AW 1995 The molecular basis of pediatric long chain 3-hydroxyyacyl-CoA dehydrogenase deficiency associated with maternal acute fatty liver of pregnancy. Proc Natl Acad Sci USA 92: 841–845
Ushikubo S, Aoyama T, Kamijo T, Wanders RJA, Rinaldo P, Vockley J, Hashimoto T 1996 Molecular characterization of mitochondrial trifunctional protein deficiency: formation of the enzyme complex is important for stabilization of both - and -subunits. Am J Hum Genet 58: 979–988
Ding J-H, Yang B-Z, Nada MA, Roe CR 1996 Improved detection of the G1528C mutation in LCHAD deficiency. Biochem Mol Med 58: 46–51
IJlst L, Ruiter JPN, Hoovers JMN, Jakobs ME, Wanders RJA 1996 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 α subunit gene. J Clin Invest 98: 1028–1033
IJlst L, Wanders RJA, Ushikubo S, Kamijo T, Hashimoto T 1994 Molecular basis of long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency: identification of the major disease-causing mutation in the α-subunit of the mitochondrial trifunctional protein. Biochim Biophys Acta 1215: 347–350
Millington DS, Kodo N, Terada N, Roe D, Chace DH 1991 The analysis of diagnostic markers of genetic disorders in human blood and urine using tandem mass spectrometry with liquid secondary ion mass spectrometry. Int J Mass Spectrom Ion Proc 111: 211–228
Millington DS, Norwood DL, Kodo N, Roe CR, Inoue F 1989 Application of fast atom bombardment with tandem mass spectrometry and liquid chromatography/mass spectrometry to the analysis of acylcarnitines in human urine, blood, and tissue. Anal Biochem 180: 331–339
Shen J-J, Matern D, Millington DS, Hillman SL, Feezor M, Bennett MJ, Qumsiyeh M, Kahler SG, Chen Y-T, Van Hove JLK 1998 Acylcarnitines produced in vitro by cultured fibroblasts of patients with long-chain 3-hydroxyacyl-coenzyme A dehydrogenase deficiency and other disorders of fatty acid oxidation. Annual SIMD Meeting, Asilomar, (abstr)
Perlick HA, Medghalchi SM, Spencer FA, Kendzior RJ, Dietz HC 1996 Mammalian orthologues of a yeast regulator of nonsense transcript stability. Proc Natl Acad Sci USA 93: 10928–10932
Rashed MS, Ozand PT, Bennett MJ, Barnard JJ, Govindaraju DR, Rinaldo P 1995 Inborn errors of metabolism diagnosed in sudden death cases by acylcarnitine analysis of postmortem bile. Clin Chem 41: 1109–1114
Tyni T, Palotie A, Viinikka L, Valanne L, Salo MK, von Döbeln U, Jackson S, Wanders R, Venizelos N, Pihko H 1997 Long-chain 3-hydroxyacyl-coenzyme A dehydrogenase deficiency with the G1528C mutation: clinical presentation of thirteen patients. J Pediatr 130: 67–76
Tyni T, Ekholm E, Pihko H 1998 Pregnancy complications are frequent in long-chain 3-hydroxyacyl-coenzyme A dehydrogenase deficiency. Am J Obstet Gynecol 178: 603–608
Roe CR 1997 Mitochondrial fatty acid oxidation disorders: chapter 45 update. In: Scriver CR, Beaudet AL, Sly WS, Valle D (eds) The Metabolic and Molecular Bases of Inherited Disease. McGraw-Hill, New York, CD-ROM
Acknowledgements
The authors thank Y.-T. Chen and Robert D. Stevens (Durham, NC) for stimulating discussion and advice, Beverly Gibson (St. Louis, MO) for expert technical assistance in molecular genetic analysis, and Andreas Schulze (Heidelberg, Germany) for provision of age-matched control blood spots.
Author information
Authors and Affiliations
Additional information
This work was partially supported by the Deutsche Forschungsgemeinschaft (MA 1964/1-1) (to Dietrich Matern).
Rights and permissions
About this article
Cite this article
Matern, D., Strauss, A., Hillman, S. et al. Diagnosis of Mitochondrial Trifunctional Protein Deficiency in a Blood Spot from the Newborn Screening Card by Tandem Mass Spectrometry and DNA Analysis. Pediatr Res 46, 45–49 (1999). https://doi.org/10.1203/00006450-199907000-00008
Received:
Accepted:
Issue date:
DOI: https://doi.org/10.1203/00006450-199907000-00008
