Skip to main content
NCBI home page
American Journal of Human Genetics logoLink to American Journal of Human Genetics
. 1987 Jan;40(1):50–59.

The French and North American phenotypes of pyruvate carboxylase deficiency, correlation with biotin containing protein by 3H-biotin incorporation, 35S-streptavidin labeling, and Northern blotting with a cloned cDNA probe.

B H Robinson, J Oei, J M Saudubray, C Marsac, K Bartlett, F Quan, R Gravel
PMCID: PMC1684008  PMID: 3101494

Abstract

Cultured skin fibroblasts from 16 patients with either French or American pyruvate carboxylase (PC) deficiency were examined for their ability to incorporate 3H-biotin into proteins. Cell extracts were also examined for the presence of biotin-containing proteins with 35S-streptavidin, immunoreactive protein with anti-PC antibody, and PC mRNA by Northern blotting with a PC cDNA probe. All the North American presentation patients showed a 3H-biotin protein, a streptavidin protein, and an anti-PC precipitable protein at 125 kilodaltons on sodium dodecyl sulfate-polyacrylamide gel electrophoresis of cellular proteins. They also showed a detectable mRNA species for PC on Northern blotting. Of the French presentation patients, five showed very low or absent 3H-biotin protein, streptavidin protein, and anti-PC precipitable protein at 125 kilodaltons. Three French presentation patients showed PC protein to be present on the basis of these techniques. Similarly, five showed either very low or absent mRNA for PC on Northern blotting whereas three gave evidence of the presence of PC-specific mRNA. Thus, whereas the North American presentation of PC deficiency is associated with the presence of a mature biotin containing protein of the correct molecular weight, the French presentation may, in some (but not in all) cases, have both absent PC protein and absent PC mRNA.

Full text

PDF
50

Images in this article

54Fig. 1
on p.54
55Fig. 2
on p.55
56Fig. 3
on p.56

Selected References

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

  1. Atkin B. M., Utter M. F., Weinberg M. B. Pyruvate carboxylase and phosphoenolpyruvate carboxykinase activity in leukocytes and fibroblasts from a patient with pyruvate carboxylase deficiency. Pediatr Res. 1979 Jan;13(1):38–43. doi: 10.1203/00006450-197901000-00009. [DOI] [PubMed] [Google Scholar]
  2. Bartlett K., Ghneim H. K., Stirk J. H., Dale G., Alberti K. G. Pyruvate carboxylase deficiency. J Inherit Metab Dis. 1984;7 (Suppl 1):74–78. doi: 10.1007/BF03047379. [DOI] [PubMed] [Google Scholar]
  3. Burri B. J., Sweetman L., Nyhan W. L. Mutant holocarboxylase synthetase: evidence for the enzyme defect in early infantile biotin-responsive multiple carboxylase deficiency. J Clin Invest. 1981 Dec;68(6):1491–1495. doi: 10.1172/JCI110402. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  5. Coude F. X., Ogier H., Marsac C., Munnich A., Charpentier C., Saudubray J. M. Secondary citrullinemia with hyperammonemia in four neonatal cases of pyruvate carboxylase deficiency. Pediatrics. 1981 Dec;68(6):914–914. [PubMed] [Google Scholar]
  6. DeVivo D. C., Haymond M. W., Leckie M. P., Bussman Y. L., McDougal D. B., Jr, Pagliara A. S. The clinical and biochemical implications of pyruvate carboxylase deficiency. J Clin Endocrinol Metab. 1977 Dec;45(6):1281–1296. doi: 10.1210/jcem-45-6-1281. [DOI] [PubMed] [Google Scholar]
  7. Freytag S. O., Collier K. J. Molecular cloning of a cDNA for human pyruvate carboxylase. Structural relationship to other biotin-containing carboxylases and regulation of mRNA content in differentiating preadipocytes. J Biol Chem. 1984 Oct 25;259(20):12831–12837. [PubMed] [Google Scholar]
  8. Haworth J. C., Robinson B. H., Perry T. L. Lactic acidosis due to pyruvate carboxylase deficiency. J Inherit Metab Dis. 1981;4(2):57–58. doi: 10.1007/BF02263589. [DOI] [PubMed] [Google Scholar]
  9. Haworth J. C., Robinson B. H., Perry T. L. Lactic acidosis due to pyruvate carboxylase deficiency. J Inherit Metab Dis. 1981;4(2):57–58. doi: 10.1007/BF02263589. [DOI] [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  12. Millis A. J., Pious D. A. Oxidative phosphorylation in mitochondria isolated from human fibroblasts. Biochim Biophys Acta. 1973 Jan 18;292(1):73–77. doi: 10.1016/0005-2728(73)90251-x. [DOI] [PubMed] [Google Scholar]
  13. Nikolau B. J., Wurtele E. S., Stumpf P. K. Use of streptavidin to detect biotin-containing proteins in plants. Anal Biochem. 1985 Sep;149(2):448–453. doi: 10.1016/0003-2697(85)90596-2. [DOI] [PubMed] [Google Scholar]
  14. Oizumi J., Shaw K. N., Giudici T. A., Carter M., Donnell G. N., Ng W. G. Neonatal pyruvate carboxylase deficiency with renal tubular acidosis and cystinuria. J Inherit Metab Dis. 1983;6(3):89–94. doi: 10.1007/BF01800731. [DOI] [PubMed] [Google Scholar]
  15. Perry T. L., Haworth J. C., Robinson B. H. Brain amino acid abnormalities in pyruvate carboxylase deficiency. J Inherit Metab Dis. 1985;8(2):63–66. doi: 10.1007/BF01801666. [DOI] [PubMed] [Google Scholar]
  16. Robinson B. H., Oei J., Saunders M., Gravel R. [3H]biotin-labeled proteins in cultured human skin fibroblasts from patients with pyruvate carboxylase deficiency. J Biol Chem. 1983 May 25;258(10):6660–6664. [PubMed] [Google Scholar]
  17. Robinson B. H., Oei J., Sherwood W. G., Applegarth D., Wong L., Haworth J., Goodyer P., Casey R., Zaleski L. A. The molecular basis for the two different clinical presentations of classical pyruvate carboxylase deficiency. Am J Hum Genet. 1984 Mar;36(2):283–294. [PMC free article] [PubMed] [Google Scholar]
  18. Robinson B. H., Taylor J., Sherwood W. G. The genetic heterogeneity of lactic acidosis: occurrence of recognizable inborn errors of metabolism in pediatric population with lactic acidosis. Pediatr Res. 1980 Aug;14(8):956–962. doi: 10.1203/00006450-198008000-00013. [DOI] [PubMed] [Google Scholar]
  19. Robinson B. H., Toone J. R., Benedict R. P., Dimmick J. E., Oei J., Applegarth D. A. Prenatal diagnosis of pyruvate carboxylase deficiency. Prenat Diagn. 1985 Jan-Feb;5(1):67–71. doi: 10.1002/pd.1970050112. [DOI] [PubMed] [Google Scholar]
  20. Saudubray J. M., Marsac C., Cathelineau C. L., Besson Leaud M., Leroux J. P. Neonatal congenital lactic acidosis with pyruvate carboxylase deficiency in two siblings. Acta Paediatr Scand. 1976 Nov;65(6):717–724. doi: 10.1111/j.1651-2227.1976.tb18009.x. [DOI] [PubMed] [Google Scholar]
  21. Saunders M. E., Sherwood W. G., Duthie M., Surh L., Gravel R. A. Evidence for a defect of holocarboxylase synthetase activity in cultured lymphoblasts from a patient with biotin-responsive multiple carboxylase deficiency. Am J Hum Genet. 1982 Jul;34(4):590–601. [PMC free article] [PubMed] [Google Scholar]
  22. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  23. Tsuchiyama A., Oyanagi K., Hirano S., Tachi N., Sogawa H., Wagatsuma K., Nakao T., Tsugawa S., Kawamura Y. A case of pyruvate carboxylase deficiency with later prenatal diagnosis of an unaffected sibling. J Inherit Metab Dis. 1983;6(3):85–88. doi: 10.1007/BF01800730. [DOI] [PubMed] [Google Scholar]
  24. Wahl G. M., Padgett R. A., Stark G. R. Gene amplification causes overproduction of the first three enzymes of UMP synthesis in N-(phosphonacetyl)-L-aspartate-resistant hamster cells. J Biol Chem. 1979 Sep 10;254(17):8679–8689. [PubMed] [Google Scholar]
  25. Wolf B., Feldman G. L. The biotin-dependent carboxylase deficiencies. Am J Hum Genet. 1982 Sep;34(5):699–716. [PMC free article] [PubMed] [Google Scholar]
  26. Wolf B., Grier R. E., Allen R. J., Goodman S. I., Kien C. L. Biotinidase deficiency: the enzymatic defect in late-onset multiple carboxylase deficiency. Clin Chim Acta. 1983 Jul 15;131(3):273–281. doi: 10.1016/0009-8981(83)90096-7. [DOI] [PubMed] [Google Scholar]
  27. Wolf B., Grier R. E., Allen R. J., Goodman S. I., Kien C. L., Parker W. D., Howell D. M., Hurst D. L. Phenotypic variation in biotinidase deficiency. J Pediatr. 1983 Aug;103(2):233–237. doi: 10.1016/s0022-3476(83)80351-5. [DOI] [PubMed] [Google Scholar]

Articles from American Journal of Human Genetics are provided here courtesy of American Society of Human Genetics

RESOURCES