Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1984 Dec 15;224(3):971–976. doi: 10.1042/bj2240971

The role of phosphoenolpyruvate carboxykinase in muscle alanine synthesis.

T N Palmer, M A Caldecourt, J P Warner, M C Sugden
PMCID: PMC1144535  PMID: 6151838

Abstract

3-Mercaptopicolinic acid (3-MPA) is reportedly a specific inhibitor of phosphoenolpyruvate (PEP) carboxykinase and has hitherto been used accordingly to elucidate the metabolic role of PEP carboxykinase in vitro and in vivo. We show that 3-MPA has multiple effects on intermediary metabolism in hemidiaphragms from 40 h-starved rats. It decreases the release of lactate + pyruvate and alanine in hemidiaphragms provided with no added substrate or with valine, leucine or isoleucine. Moreover, irrespective of the substrate provided (none, valine, leucine, isoleucine, glucose, acetate, oleate), 3-MPA decreases the [lactate]/[pyruvate] ratio. 3-MPA is without effect on 14CO2 production from [U-14C]valine, [1-14C]valine, [1-14C]leucine, [U-14C]isoleucine or [1-14C]oleate, but stimulates 14CO2 production from [U-14C]glucose and [1-14C]pyruvate and inhibits 14CO2 production from [1-14C]acetate. Glycolytic flux (measured as 3H2O formation from [5-3H]glucose) is stimulated by 3-MPA. It is concluded that 3-MPA has site(s) of actions other than PEP carboxykinase and that the putative role of PEP carboxykinase in alanine synthesis de novo in skeletal muscle from tricarboxylic acid-cycle intermediates and related amino acids requires reappraisal.

Full text

PDF
975

Selected References

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

  1. Ashcroft S. J., Weerasinghe L. C., Bassett J. M., Randle P. J. The pentose cycle and insulin release in mouse pancreatic islets. Biochem J. 1972 Feb;126(3):525–532. doi: 10.1042/bj1260525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blackshear P. J., Holloway P. A., Aberti K. G. The effects of inhibition of gluconeogenesis on ketogenesis in starved and diabetic rats. Biochem J. 1975 Jun;148(3):353–362. doi: 10.1042/bj1480353b. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chang T. W., Goldberg A. L. The origin of alanine produced in skeletal muscle. J Biol Chem. 1978 May 25;253(10):3677–3684. [PubMed] [Google Scholar]
  4. DiTullio N. W., Berkoff C. E., Blank B., Kostos V., Stack E. J., Saunders H. L. 3-mercaptopicolinic acid, an inhibitor of gluconeogenesis. Biochem J. 1974 Mar;138(3):387–394. doi: 10.1042/bj1380387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Duff D. A., Snell K. Limitations of commonly used spectrophotometric assay methods for phosphoenolypyruvate carboxykinase activity in crude extracts of muscle. Biochem J. 1982 Jul 15;206(1):147–152. doi: 10.1042/bj2060147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Felig P. Amino acid metabolism in man. Annu Rev Biochem. 1975;44:933–955. doi: 10.1146/annurev.bi.44.070175.004441. [DOI] [PubMed] [Google Scholar]
  7. Garber A. J., Karl I. E., Kipnis D. M. Alanine and glutamine synthesis and release from skeletal muscle. IV. beta-Adrenergic inhibition of amino acid release. J Biol Chem. 1976 Feb 10;251(3):851–857. [PubMed] [Google Scholar]
  8. Goldstein L., Newsholme E. A. The formation of alanine from amino acids in diaphragm muscle of the rat. Biochem J. 1976 Feb 15;154(2):555–558. doi: 10.1042/bj1540555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Goodman M. N. Effect of 3-mercaptopicolinic acid on gluconeogenesis and gluconeogenic metabolite concentrations in the isolated perfused rat liver. Biochem J. 1975 Jul;150(1):137–139. doi: 10.1042/bj1500137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kostos V., DiTullio N. W., Rush J., Cieslinski L., Saunders H. L. The effect of 3-mercaptopicolinic acid on phosphoenolpyruvate carboxykinase (GTP) in the rat and guinea pig. Arch Biochem Biophys. 1975 Dec;171(2):459–465. doi: 10.1016/0003-9861(75)90054-5. [DOI] [PubMed] [Google Scholar]
  11. Nordlie R. C., Alvares F. L., Sukalski K. A. Stimulation by 3-mercaptopicolinate of net glucose uptake by perfused livers from diabetic rats. Biochim Biophys Acta. 1982 Nov 24;719(2):244–250. doi: 10.1016/0304-4165(82)90095-2. [DOI] [PubMed] [Google Scholar]
  12. Palmer T. N., Caldecourt M. A., Slavin J. P. Pyruvate kinase and alanine synthesis in skeletal muscle. Biosci Rep. 1982 Nov;2(11):941–948. doi: 10.1007/BF01114901. [DOI] [PubMed] [Google Scholar]
  13. Palmer T. N., Caldecourt M. A., Sugden M. C. Adrenergic inhibition of branched-chain 2-oxo acid dehydrogenase in rat diaphragm muscle in vitro. Biochem J. 1983 Oct 15;216(1):63–70. doi: 10.1042/bj2160063. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Poston J. M. The relative carbon flux through the alpha- and the beta-keto pathways of leucine metabolism. J Biol Chem. 1984 Feb 25;259(4):2059–2061. [PubMed] [Google Scholar]
  15. Robinson B. H., Oei J. 3-Mercaptopicolinic acid, a preferential inhibitor of the cytosolic phosphoenolpyruvate carboxykinase. FEBS Lett. 1975 Oct 15;58(1):12–15. doi: 10.1016/0014-5793(75)80214-6. [DOI] [PubMed] [Google Scholar]
  16. Ruderman N. B. Muscle amino acid metabolism and gluconeogenesis. Annu Rev Med. 1975;26:245–258. doi: 10.1146/annurev.me.26.020175.001333. [DOI] [PubMed] [Google Scholar]
  17. Snell K., Duff D. A. The hepato-muscular metabolic axis and gluconeogenesis. Prog Clin Biol Res. 1982;102(Pt 100):279–291. [PubMed] [Google Scholar]
  18. Snell K., Duff D. A. The release of alanine by rat diaphragm muscle in vitro. Biochem J. 1977 Feb 15;162(2):399–403. doi: 10.1042/bj1620399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Snell K. Muscle alanine synthesis and hepatic gluconeogenesis. Biochem Soc Trans. 1980 Apr;8(2):205–213. doi: 10.1042/bst0080205. [DOI] [PubMed] [Google Scholar]

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

RESOURCES