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Accumulation of methylmalonic acid caused by vitamin B12-deficiency disrupts normal cellular metabolism in rat liver

Published online by Cambridge University Press:  09 March 2007

Shigeki Toyoshima ,
Fumio Watanabe ,
Hisako Saido ,
Ewa H. Pezacka ,
Donald W. Jacobsens ,
Kazutaka Miyatake  and
Yoshihisa Nakano
Shigeki Toyoshima
Affiliation:
Department of Applied Biological Chemistry, Osaka Prefecture University, Sakai, Osaka 593, Japan
Fumio Watanabe
Affiliation:
Department of Foods and Nutrition, Kochi Women's University, Kochi 780, Japan
Hisako Saido
Affiliation:
Department of Applied Biological Chemistry, Osaka Prefecture University, Sakai, Osaka 593, Japan
Ewa H. Pezacka
Affiliation:
Departments of Cell Biology and Clinical Pathology, The Cleveland Clinic Foundation, Cleveland, Ohio 44195, USA
Donald W. Jacobsens
Affiliation:
Departments of Cell Biology and Clinical Pathology, The Cleveland Clinic Foundation, Cleveland, Ohio 44195, USA
Kazutaka Miyatake
Affiliation:
Department of Applied Biological Chemistry, Osaka Prefecture University, Sakai, Osaka 593, Japan
Yoshihisa Nakano
Affiliation:
Department of Applied Biological Chemistry, Osaka Prefecture University, Sakai, Osaka 593, Japan
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Abstract

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To clarify the relationship between intracellular concentrations of methylmalonic acid and metabolic and growth inhibition in vitamin B12-deficient rats, hepatic methylmalonic acidlevels were assayed and inhibition of glucose and glutamic acid metabolism by methylmalonic acid was studied in isolated hepatocytes. Vitamin B12-deficient rats (14 weeks old) excreted more urinary methylmalonic acid and had lower body weights than the control rats. Hepatic methylmalonic acid levels (3·6 (SD 1·30)–5·3 (SD 0·51) µmol/g tissue; 7·9 (SD 2·90)–11·8 (SD 1·14) mM) were increased and correlated with the extent of the growth retardation during vitamin B12-deficiency. Isolated hepatocytes and mitochondria from normally fed rats were labelled with [14C(U)]glucose and [14C(U)]glutamic acid respectively, in the presence or absence of 5mM-methylmalonic acid. Although methylmalonic acid did not affect the incorporation of 14C into protein and organic acid fractions in the hepatocytes, it inhibited 14CO2 formation (an index of glucose oxidation by the Krebs cycle) by 25% and incorporation of 14C into the amino acid fractionby 30%. In the mitochondria, methylmalonic acid inhibited 14CO2, formation (indicating glutamic acid oxidation by the Krebs cycle) by 70%, but not the incorporation of 14C into the protein fraction. The incorporation of 14C into the organic acid fraction was significantly stimulated by the addition of methylmalonic acid. These results indicate that the unusual accumulation of methylmalonic acid caused by vitamin B12-deficiency disrupts normal glucose and glutamic acid metabolism in rat liver, probably by inhibiting the Krebs cycle.

Keywords

Vitamin B12Methylmalonic acidLiver
Type
Cobalamin deficiency
Information
British Journal of Nutrition , Volume 75 , Issue 6 , June 1996 , pp. 929 - 938
Copyright
Copyright © The Nutrition Society 1996

References

REFERENCES

Arinze, I.J., Waters, D. & Donaldson, M. K. (1979). Effect of methylmalonic acid on gluconeogenesis in isolated rat and guinea-pig hepatocytes. Biochemical Journal 184, 717–719.CrossRefGoogle Scholar
Berry, M. N. & Friend, D. S. (1969). High-yield preparation of isolated rat liver parenchymal cells. A biochemical and fine Structure study, Journal of cell Biology 43, 506–520.CrossRefGoogle Scholar
Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248–254CrossRefGoogle ScholarPubMed
Brass, E. P., Fennessey, P. V. & Miller, L. V. (1986). Inhibition of oxidative metabolism by propionic acid and its reversal by carnitine in isolated rat hepatocytes. Biochemical Journal 236,131–136CrossRefGoogle ScholarPubMed
Brass, E. P. & Stabler, S. P. (1988). Carnitine metabolism in the vitamin B-12-deficient rat. Biochemical Journal 155, 153–159.CrossRefGoogle Scholar
Brass, E. P., Tahiliani, A. G., Allen, R. H. & Stabler, S. P. (1990). Coenzyme A metabolismin vitamin B-12- deficient rats. Journal of Nutrition 120, 290–297.CrossRefGoogle Scholar
Cardinale, G. J., Dreyfus, P. M., Auld, P. & Abeles, R. H. (1969). Experimental vitamin B12deficiency its effect on tissue vitamin B12coenzyme levels and on the metabolism of methylmalonyl-CoA. Archives of Biochemistry and Biophysics 131, 92–99.CrossRefGoogle Scholar
Dryden, L. P. & Hartman, A. M. (1966). Effects of vitamin B-12 on the weights of certain organs in the rat. Journal of Nutrition 90, 377–381.CrossRefGoogle ScholarPubMed
Fchling, C., Jagerstad, M., Akesson, B., Axelsson, J. & Brun, A. (1978). Effects of vitamin B12 deficiency on lipid metabolism of the rat liver and nervous system. British Journal of Nutrition 39, 501–513.CrossRefGoogle Scholar
Frenkel, E. P., Kitchens, R. L. & Johnston, J. H. (1973). The effect of vitamin B12 deprivation on the enzymes of fatty acid synthesis. Journal of Biological Chemistry 248, 7540–7546.CrossRefGoogle ScholarPubMed
Frenkel, E. P., Kitchens, R. L., Hersh, L. B. & Frenkel, R. (1974). Effect of vitamin B12 deprivation on the in vivo levels of coenzyme A intermediates associated with propionate metabolism. Journal of Biological Chemistry 249, 6984–6991.CrossRefGoogle Scholar
Frenkel, E. P., Mukherjee, A., Hackenbrock, C. R. & Srere, P. A. (1976). Biochemical and ultrastructural hepatic changes during vitamin B12 deficiency in animals and man. Journal of Biological Chemistry 251, 2147–2154.CrossRefGoogle Scholar
Frenkel, E. P., Prough, R. & Kitchens, R. L. (1980). Measurement of tissue vitamin B12, by radioisotopic competitive inhibition assay and a quantitation of tissue cobalamin fractions. Methods of Enzymology 67,31–40.CrossRefGoogle Scholar
Halperin, M. L., Schiller, C. M. & Fritz, I. B. (1971). The inhibition by methylmalonic acid of malate transport by the dicarboxylate carrier in rat liver mitochondria. A possible explanation for hypoglycemia in methylmalonic aciduria. Journal of Clinical Investigation 50 2276–2282.CrossRefGoogle Scholar
Jacobsen, D. W., Gatautis, V. J. & Green, R. (1989). Determination of plasma homocysteine by high-performance liquid chromatography with fluorescence detection. Analytical Biochemistry 178, 208–214.CrossRefGoogle ScholarPubMed
Jacobsen, D. W., Gatautis, V. J., Green, R., Robinson, K., Savon, S. R., Secic, M.Ji, J., Otto, J. M. & Taylor, L.M. Jr(1994). Rapid HPLC determination of total homocysteine and other thiols in serum and plasma sex differences and correlation with cobalamin and folate levels in normal subjects. Clinical Chemistry 40, 873–881.CrossRefGoogle Scholar
Kennedy, D. G., Blanchflower, W. J., Scott, J. M., Weir, D. G., Molloy, A. M., Kennedy, S. & Young, P. B. (1992). Cobalt-vitamin B-12 deficiency decreases methionine synthase activity and phospholipid methylation in sheep. Journal of Nutrition 122, 1384–1390.CrossRefGoogle ScholarPubMed
Kennedy, D. G., Cannavan, A., Molloy, A., O'Harte, F. O., Taylor, S. M., Kennedy, S. & Blanchflower, W. J.(1990). Methylmalonyl-CoA mutase (EC 5.4.99.2) and methionine synthetase (EC 2.1. 1.13) in the tissues of cobalt-vitamin B12 deficient sheep. British Journal of Nutrition 64, 721–732.CrossRefGoogle ScholarPubMed
McCully, K. S. (1992). Homocysteinuria, arteriosclerosis, methylmalonic aciduria, and methyltransferase deficiency a key case revisited. Nutrition Reviews 50, 7–12.CrossRefGoogle Scholar
Martin-Requero, A., Corkey, B. E., Cerdan, S., Walajtys-Rode, E., Parrilla, R. L. & Williamson, J. R. (1973).Interactions between α-ketoisovalerate metabolism and the pathways of gluconeogenesis and urea synthesis in isolated hepatocytes. Journal of Biological Chemistry 258, 3673–3681.CrossRefGoogle Scholar
Metz, J. (1992). Cobalamin deficiency and the pathogenesis of nervous system disease. Annual Review of Nutrition 12, 59–79.CrossRefGoogle Scholar
Moelby, L., Rasmussen, K., Jensen, M. K. & Pedersen, K. O. (1990). The relationship betweenclinically confirmed cobalamin deficiency and serum methylmalonic acid. Journal of Internal Medicine 228, 373–378.CrossRefGoogle Scholar
National Research Council (1985). Guide for the Care and Use of Laboratory Animals. Publication no. 85–23 (revised). Bethesda: National Institutes of Health.Google Scholar
Siess, E. A., Brocks, D. G. &Wieland, O. H. (1982). Subcellular distribution of adenine nucleotides and metabolites of tricarboxylate cycle and gluconeogenesis in hepatocytes. In Metabolic Compartmentation, pp.235–257 [Sies, H., editor]. London: Academic Press.Google Scholar
Toyoshima, S., Saido, S., Watanabe, F., Miyatake, K. & Nakano, Y. (1994). Assay for urinary methylmalonic acid by high-pressure liquid chromatography. Bioscience, Biotechnology, and Biochemistry 58, 1882–1883.CrossRefGoogle Scholar
Watanabe, F., Nakano, Y., Stupperich, E., Ushikoshi, K., Ushikoshi, S., Ushikoshi, I. & Kitaoka, S. (1993). A radioisotope dilution method for quantitation of total vitamin B12 in biological samples using isolated Euglena pellicle fragments as a solid-phase vitamin B12-binding material. Analytical Chemistry 65, 657–659.CrossRefGoogle Scholar
Watanabe, F., Nakano, Y., Tachikake, N., Saido, H., Tamura, Y. & Yamanaka, H. (1991). Vitamin B-12 deficiency increases the specific activities of rat liver NADH- and NADPH-linked aquacobalamin reductase isozymes involved in coenzyme synthesis. Journal of Nutrition 121, 1948–1954.CrossRefGoogle ScholarPubMed
Watanabe, F., Saido, H., Toyoshima, S., Tamura, Y. & Nakano, Y. (1994). Feeding of vitaminB12 rapidly increases the specific activity of hepatic methylmalonyl-CoA mutase in vitamin B12deficient rats. Bioscience, Biotechnology, and Biochemistry 58, 556–557.CrossRefGoogle Scholar
Weidemann, M. J., Hems, R., Williams, D. L., Spray, G. H. & Krebs, H. A. (1970). Gluconeogenesis from propionate kidney and liver of the vitamin B12-deficient rat. Biochemical Journal 117, 177–181.CrossRefGoogle ScholarPubMed
Williams, D. L. & Spray, G. H. (1971). Metabolic effects of propionate in normal and vitamin B12-deficient rats. Biochemical Journal 124, 501–507.CrossRefGoogle Scholar