Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1996 Jul 15;317(Pt 2):613–619. doi: 10.1042/bj3170613

Measurement of the turnover of glycogen phosphorylase by GC/MS using stable isotope derivatives of pyridoxine (vitamin B6).

R J Beynon 1, D M Leyland 1, R P Evershed 1, R H Edwards 1, S P Coburn 1
PMCID: PMC1217530  PMID: 8713093

Abstract

The majority of vitamin B6 in the body is in skeletal muscle, bound as the cofactor pyridoxal 5'-phosphate to one abundant protein, glycogen phosphorylase. Previous work has established that radiolabelled vitamin B6 can be used as a turnover label for glycogen phosphorylase. In this study, a stable isotope derivative of pyridoxine ¿dideuterated pyridoxine; 3-hydroxy-4-(hydroxymethyl) -5-[hydroxymethyl-2H2]-2-methylpyridine¿ ([2H2]PN) has been used as a metabolic tracer to study the kinetics of labelling of the body pools of vitamin B6 in mice. A non-invasive method was developed in which the isotope abundance of the urinary excretory product of vitamin B6 metabolism, 4-pyridoxic acid, was analysed by GC/MS. The change in isotope abundance of urinary 4-pyridoxic acid following administration of [2H2]PN reflects the kinetics of labelling of the body pools of vitamin B6, and yields, non-invasively, the rate of degradation of glycogen phosphorylase.

Full Text

The Full Text of this article is available as a PDF (479.4 KB).

Selected References

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

  1. Bartram C., Edwards R. H., Beynon R. J. McArdle's disease-muscle glycogen phosphorylase deficiency. Biochim Biophys Acta. 1995 Aug 15;1272(1):1–13. doi: 10.1016/0925-4439(95)00060-h. [DOI] [PubMed] [Google Scholar]
  2. Bartram C., Edwards R. H., Clague J., Beynon R. J. McArdle's disease: a nonsense mutation in exon 1 of the muscle glycogen phosphorylase gene explains some but not all cases. Hum Mol Genet. 1993 Aug;2(8):1291–1293. doi: 10.1093/hmg/2.8.1291. [DOI] [PubMed] [Google Scholar]
  3. Bartram C., Edwards R. H., Clague J., Beynon R. J. McArdle's disease: a rare frameshift mutation in exon 1 of the muscle glycogen phosphorylase gene. Biochim Biophys Acta. 1994 Jul 18;1226(3):341–343. doi: 10.1016/0925-4439(94)90047-7. [DOI] [PubMed] [Google Scholar]
  4. Beynon R. J., Bartram C., Hopkins P., Toescu V., Gibson H., Phoenix J., Edwards R. H. McArdle's disease: molecular genetics and metabolic consequences of the phenotype. Muscle Nerve Suppl. 1995;3:S18–S22. doi: 10.1002/mus.880181406. [DOI] [PubMed] [Google Scholar]
  5. Beynon R. J., Fairhurst D., Cookson E. J. Turnover of skeletal muscle glycogen phosphorylase. Biomed Biochim Acta. 1986;45(11-12):1619–1625. [PubMed] [Google Scholar]
  6. Black A. L., Guirard B. M., Snell E. E. The behavior of muscle phosphorylase as a reservoir for vitamin B6 in the rat. J Nutr. 1978 Apr;108(4):670–677. doi: 10.1093/jn/108.4.670. [DOI] [PubMed] [Google Scholar]
  7. Bode W., Mocking J. A., van den Berg H. Influence of age and sex on vitamin B-6 vitamer distribution and on vitamin B-6 metabolizing enzymes in Wistar rats. J Nutr. 1991 Mar;121(3):318–329. doi: 10.1093/jn/121.3.318. [DOI] [PubMed] [Google Scholar]
  8. Coburn S. P. Application of models to the determination of nutrient requirements: an overview. J Nutr. 1992 Mar;122(3 Suppl):687–689. doi: 10.1093/jn/122.suppl_3.687. [DOI] [PubMed] [Google Scholar]
  9. Coburn S. P., Lewis D. L., Fink W. J., Mahuren J. D., Schaltenbrand W. E., Costill D. L. Human vitamin B-6 pools estimated through muscle biopsies. Am J Clin Nutr. 1988 Aug;48(2):291–294. doi: 10.1093/ajcn/48.2.291. [DOI] [PubMed] [Google Scholar]
  10. Coburn S. P. Location and turnover of vitamin B6 pools and vitamin B6 requirements of humans. Ann N Y Acad Sci. 1990;585:76–85. doi: 10.1111/j.1749-6632.1990.tb28043.x. [DOI] [PubMed] [Google Scholar]
  11. Coburn S. P., Mahuren J. D., Erbelding W. F., Townsend D. W., Hachey D. L., Klein P. D. Measurement of vitamin B6 kinetics in vivo using chronic administration of labelled pyridoxine. Prog Clin Biol Res. 1984;144A:43–54. [PubMed] [Google Scholar]
  12. Coburn S. P., Mahuren J. D., Kennedy M. S., Schaltenbrand W. E., Sampson D. A., O'Connor D. K., Snyder D. L., Wostmann B. S. B6 vitamer content of rat tissues measured by isotope tracer and chromatographic methods. Biofactors. 1988 Dec;1(4):307–312. [PubMed] [Google Scholar]
  13. Coburn S. P., Mahuren J. D., Wostmann B. S., Snyder D. L., Townsend D. W. Role of intestinal microflora in the metabolism of vitamin B-6 and 4'-deoxypyridoxine examined using germfree guinea pigs and rats. J Nutr. 1989 Feb;119(2):181–188. doi: 10.1093/jn/119.2.181. [DOI] [PubMed] [Google Scholar]
  14. Coburn S. P., Townsend D. W. Modelling vitamin B6 metabolism in rodents (review). In Vivo. 1989 May-Jun;3(3):215–223. [PubMed] [Google Scholar]
  15. Coburn S. P., Ziegler P. J., Costill D. L., Mahuren J. D., Fink W. J., Schaltenbrand W. E., Pauly T. A., Pearson D. R., Conn P. S., Guilarte T. R. Response of vitamin B-6 content of muscle to changes in vitamin B-6 intake in men. Am J Clin Nutr. 1991 Jun;53(6):1436–1442. doi: 10.1093/ajcn/53.6.1436. [DOI] [PubMed] [Google Scholar]
  16. Cookson E. J., Beynon R. J. Further evaluation of cofactor as a turnover label for glycogen phosphorylase. Int J Biochem. 1989;21(9):975–982. doi: 10.1016/0020-711x(89)90229-2. [DOI] [PubMed] [Google Scholar]
  17. Flannery A. V., Easterby J. S., Beynon R. J. Turnover of glycogen phosphorylase in the pectoralis muscle of broiler and layer chickens. Biochem J. 1992 Sep 15;286(Pt 3):915–922. doi: 10.1042/bj2860915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Johansson S., Lindstedt S., Register U. Metabolism of labeled pyridoxine in the rat. Am J Physiol. 1966 May;210(5):1086–1095. doi: 10.1152/ajplegacy.1966.210.5.1086. [DOI] [PubMed] [Google Scholar]
  19. Johansson S., Lindstedt S., Register U., Wadström L. Studies on the metabolism of labeled pyridoxine in man. Am J Clin Nutr. 1966 Mar;18(3):185–196. doi: 10.1093/ajcn/18.3.185. [DOI] [PubMed] [Google Scholar]
  20. KREBS E. G., FISCHER E. H. PHOSPHORYLASE AND RELATED ENZYMES OF GLYCOGEN METABOLISM. Vitam Horm. 1964;22:399–410. doi: 10.1016/s0083-6729(08)60345-3. [DOI] [PubMed] [Google Scholar]
  21. Leyland D. M., Beynon R. J. The expression of glycogen phosphorylase in normal and dystrophic muscle. Biochem J. 1991 Aug 15;278(Pt 1):113–117. doi: 10.1042/bj2780113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Leyland D. M., Evershed R. P., Edwards R. H., Beynon R. J. Application of gas chromatography-mass spectrometry with selected ion monitoring to the urinanalysis of 4-pyridoxic acid. J Chromatogr. 1992 Oct 23;581(2):179–185. doi: 10.1016/0378-4347(92)80270-z. [DOI] [PubMed] [Google Scholar]
  23. Leyland D. M., Turner P. C., Beynon R. J. Effect of denervation on the expression of glycogen phosphorylase in mouse skeletal muscle. Biochem J. 1990 Nov 15;272(1):231–237. doi: 10.1042/bj2720231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lyne A., Boston R., Pettigrew K., Zech L. EMSA: a SAAM service for the estimation of population parameters based on model fits to identically replicated experiments. Comput Methods Programs Biomed. 1992 Jul;38(2-3):117–151. doi: 10.1016/0169-2607(92)90082-i. [DOI] [PubMed] [Google Scholar]
  25. McConchie S. M., Coakley J., Edwards R. H., Beynon R. J. Molecular heterogeneity in McArdle's disease. Biochim Biophys Acta. 1990 Nov 14;1096(1):26–32. doi: 10.1016/0925-4439(90)90008-d. [DOI] [PubMed] [Google Scholar]
  26. Mommaerts W. F., Illingworth B., Pearson C. M., Guillory R. J., Seraydarian K. A FUNCTIONAL DISORDER OF MUSCLE ASSOCIATED WITH THE ABSENCE OF PHOSPHORYLASE. Proc Natl Acad Sci U S A. 1959 Jun;45(6):791–797. doi: 10.1073/pnas.45.6.791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. SCHMID R., MAHLER R. Chronic progressive myopathy with myoglobinuria: demonstration of a glycogenolytic defect in the muscle. J Clin Invest. 1959 Nov;38:2044–2058. doi: 10.1172/JCI103983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tsujino S., Shanske S., DiMauro S. Molecular genetic heterogeneity of myophosphorylase deficiency (McArdle's disease). N Engl J Med. 1993 Jul 22;329(4):241–245. doi: 10.1056/NEJM199307223290404. [DOI] [PubMed] [Google Scholar]

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

RESOURCES