Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1990 Mar 15;266(3):809–815.

Effect of hydroxycobalamin[c-lactam] on propionate and carnitine metabolism in the rat.

E P Brass 1, R H Allen 1, L J Ruff 1, S P Stabler 1
PMCID: PMC1131211  PMID: 2327967

Abstract

The administration in vivo of the cobalamin analogue hydroxycobalamin[c-lactam] inhibits hepatic L-methylmalonyl-CoA mutase activity. The current studies characterize in vivo and in vitro the hydroxycobalamin[c-lactam]-treated rat as a model of disordered propionate and methylmalonic acid metabolism. Treatment of rats with hydroxycobalamin[c-lactam] (2 micrograms/h by osmotic minipump) increased urinary methylmalonic acid excretion from 0.55 mumol/day to 390 mumol/day after 2 weeks. Hydroxycobalamin[c-lactam] treatment was associated with increased urinary propionylcarnitine excretion and increased short-chain acylcarnitine concentrations in plasma and liver. Hepatocytes isolated from cobalamin-analogue-treated rats metabolized propionate (1.0 mM) to CO2 and glucose at rates which were only 18% and 1% respectively of those observed in hepatocytes from control (saline-treated) rats. In contrast, rates of pyruvate and palmitate oxidation were higher than control in hepatocytes from the hydroxycobalamin[c-lactam]-treated rats. In hepatocytes from hydroxycobalamin[c-lactam]-treated rats, propionylcarnitine was the dominant product generated from propionate when carnitine (10 mM) was present. The addition of carnitine thus resulted in a 4-fold increase in total propionate utilization under these conditions. Hepatocytes from hydroxycobalamin[c-lactam]-treated rats were more sensitive than control hepatocytes to inhibition of palmitate oxidation by propionate. This inhibition of palmitate oxidation was partially reversed by addition of carnitine. Thus hydroxycobalamin[c-lactam] treatment in vivo rapidly causes a severe defect in propionate metabolism. The consequences of this metabolic defect in vivo and in vitro are those predicted on the basis of propionyl-CoA and methylmalonyl-CoA accumulation. The cobalamin-analogue-treated rat provides a useful model for studying metabolism under conditions of a metabolic defect causing acyl-CoA accretion.

Full text

PDF
809

Selected References

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

  1. Berry M. N., Friend D. S. High-yield preparation of isolated rat liver parenchymal cells: a biochemical and fine structural study. J Cell Biol. 1969 Dec;43(3):506–520. doi: 10.1083/jcb.43.3.506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bieber L. L., Emaus R., Valkner K., Farrell S. Possible functions of short-chain and medium-chain carnitine acyltransferases. Fed Proc. 1982 Oct;41(12):2858–2862. [PubMed] [Google Scholar]
  3. Binder M., Kolhouse J. F., Van Horne K. C., Allen R. H. High-pressure liquid chromatography of cobalamins and cobalamin analogs. Anal Biochem. 1982 Sep 15;125(2):253–258. doi: 10.1016/0003-2697(82)90003-3. [DOI] [PubMed] [Google Scholar]
  4. Brass E. P., Beyerinck R. A. Effects of propionate and carnitine on the hepatic oxidation of short- and medium-chain-length fatty acids. Biochem J. 1988 Mar 15;250(3):819–825. doi: 10.1042/bj2500819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brass E. P., Beyerinck R. A. Interactions of propionate and carnitine metabolism in isolated rat hepatocytes. Metabolism. 1987 Aug;36(8):781–787. doi: 10.1016/0026-0495(87)90117-x. [DOI] [PubMed] [Google Scholar]
  6. Brass E. P. Effect of alpha-ketobutyrate on palmitic acid and pyruvate metabolism in isolated rat hepatocytes. Biochim Biophys Acta. 1986 Aug 29;888(1):18–24. doi: 10.1016/0167-4889(86)90065-0. [DOI] [PubMed] [Google Scholar]
  7. Brass E. P., Fennessey P. V., Miller L. V. Inhibition of oxidative metabolism by propionic acid and its reversal by carnitine in isolated rat hepatocytes. Biochem J. 1986 May 15;236(1):131–136. doi: 10.1042/bj2360131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brass E. P., Garrity M. J., Robertson R. P. Inhibition of glucagon-stimulated hepatic glycogenolysis by E-series prostaglandins. FEBS Lett. 1984 Apr 24;169(2):293–296. doi: 10.1016/0014-5793(84)80336-1. [DOI] [PubMed] [Google Scholar]
  9. Brass E. P., Hoppel C. L. Carnitine metabolism in the fasting rat. J Biol Chem. 1978 Apr 25;253(8):2688–2693. [PubMed] [Google Scholar]
  10. Brass E. P., Hoppel C. L. Relationship between acid-soluble carnitine and coenzyme A pools in vivo. Biochem J. 1980 Sep 15;190(3):495–504. doi: 10.1042/bj1900495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Brass E. P., Ruff L. J. Effect of carnitine on propionate metabolism in the vitamin B-12--deficient rat. J Nutr. 1989 Aug;119(8):1196–1202. doi: 10.1093/jn/119.8.1196. [DOI] [PubMed] [Google Scholar]
  12. Brass E. P., Stabler S. P. Carnitine metabolism in the vitamin B-12-deficient rat. Biochem J. 1988 Oct 1;255(1):153–159. doi: 10.1042/bj2550153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Burton W. C., Frenkel E. P. Effect of vitamin B-12 deficiency on the hepatic tissue concentration of acyl carnitines. Biochim Biophys Acta. 1975 Aug 25;398(2):217–223. doi: 10.1016/0005-2760(75)90137-x. [DOI] [PubMed] [Google Scholar]
  14. Cardinale G. J., Dreyfus P. M., Auld P., Abeles R. H. Experimental vitamin B12 deficiency: its effect on tissue vitamin B12-coenzyme levels and on the metabolism of methylmalonyl-CoA. Arch Biochem Biophys. 1969 Apr;131(1):92–99. doi: 10.1016/0003-9861(69)90108-8. [DOI] [PubMed] [Google Scholar]
  15. Cederblad G., Lindstedt S. A method for the determination of carnitine in the picomole range. Clin Chim Acta. 1972 Mar;37:235–243. doi: 10.1016/0009-8981(72)90438-x. [DOI] [PubMed] [Google Scholar]
  16. Ciman M., Rossi C. R., Siliprandi N. On the mechanism of the antiketogenic action of propionate and succinate in isolated rat liver mitochondria. FEBS Lett. 1972 Apr 15;22(1):8–10. doi: 10.1016/0014-5793(72)80205-9. [DOI] [PubMed] [Google Scholar]
  17. Cooper B. A., Rosenblatt D. S. Inherited defects of vitamin B12 metabolism. Annu Rev Nutr. 1987;7:291–320. doi: 10.1146/annurev.nu.07.070187.001451. [DOI] [PubMed] [Google Scholar]
  18. Frenkel E. P., Kitchens R. L., Hersh L. B., Frenkel R. Effect of vitamin B12 deprivation on the in vivo levels of coenzyme A intermediates associated with propionate metabolism. J Biol Chem. 1974 Nov 10;249(21):6984–6991. [PubMed] [Google Scholar]
  19. Frenkel E. P., White J. D. Characterization of an animal model of vitamin B12 deprivation. Lab Invest. 1973 Dec;29(6):614–619. [PubMed] [Google Scholar]
  20. Glasgow A. M., Chase H. P. Effect of propionic acid on fatty acid oxidation and ureagenesis. Pediatr Res. 1976 Jul;10(7):683–686. doi: 10.1203/00006450-197607000-00010. [DOI] [PubMed] [Google Scholar]
  21. Lysiak W., Lilly K., DiLisa F., Toth P. P., Bieber L. L. Quantitation of the effect of L-carnitine on the levels of acid-soluble short-chain acyl-CoA and CoASH in rat heart and liver mitochondria. J Biol Chem. 1988 Jan 25;263(3):1151–1156. [PubMed] [Google Scholar]
  22. Lysiak W., Toth P. P., Suelter C. H., Bieber L. L. Quantitation of the efflux of acylcarnitines from rat heart, brain, and liver mitochondria. J Biol Chem. 1986 Oct 15;261(29):13698–13703. [PubMed] [Google Scholar]
  23. MARQUIS N. R., FRITZ I. B. ENZYMOLOGICAL DETERMINATION OF FREE CARNITINE CONCENTRATIONS IN RAT TISSUES. J Lipid Res. 1964 Apr;5:184–187. [PubMed] [Google Scholar]
  24. Marcell P. D., Stabler S. P., Podell E. R., Allen R. H. Quantitation of methylmalonic acid and other dicarboxylic acids in normal serum and urine using capillary gas chromatography-mass spectrometry. Anal Biochem. 1985 Oct;150(1):58–66. doi: 10.1016/0003-2697(85)90440-3. [DOI] [PubMed] [Google Scholar]
  25. Martin-Requero A., Corkey B. E., Cerdan S., Walajtys-Rode E., Parrilla R. L., Williamson J. R. Interactions between alpha-ketoisovalerate metabolism and the pathways of gluconeogenesis and urea synthesis in isolated hepatocytes. J Biol Chem. 1983 Mar 25;258(6):3673–3681. [PubMed] [Google Scholar]
  26. McGarry J. D., Foster D. W. Regulation of hepatic fatty acid oxidation and ketone body production. Annu Rev Biochem. 1980;49:395–420. doi: 10.1146/annurev.bi.49.070180.002143. [DOI] [PubMed] [Google Scholar]
  27. Mills S. E., Foster D. W., McGarry J. D. Interaction of malonyl-CoA and related compounds with mitochondria from different rat tissues. Relationship between ligand binding and inhibition of carnitine palmitoyltransferase I. Biochem J. 1983 Jul 15;214(1):83–91. doi: 10.1042/bj2140083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Stabler S. P., Marcell P. D., Podell E. R., Allen R. H., Lindenbaum J. Assay of methylmalonic acid in the serum of patients with cobalamin deficiency using capillary gas chromatography-mass spectrometry. J Clin Invest. 1986 May;77(5):1606–1612. doi: 10.1172/JCI112476. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wałajtys-Rode E., Coll K. E., Williamson J. R. Effects of branched chain alpha-ketoacids on the metabolism of isolated rat liver cells. II. Interactions with gluconeogenesis and urea synthesis. J Biol Chem. 1979 Nov 25;254(22):11521–11529. [PubMed] [Google Scholar]
  30. Weidemann M. J., Hems R., Williams D. L., Spray G. H., Krebs H. A. Gluconeogenesis from propionate in kidney and liver of the vitamin B12-deficient rat. Biochem J. 1970 Mar;117(1):177–181. doi: 10.1042/bj1170177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Williams D. L., Spray G. H., Newman G. E., O'Brien J. R. Dietary depletion of vitamin B12 and the excretion of methylmalonic acid in the rat. Br J Nutr. 1969 Jun;23(2):343–352. doi: 10.1079/bjn19690041. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES