Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1989 Oct 1;263(1):231–241. doi: 10.1042/bj2630231

A 13C-n.m.r. investigation of the metabolism of amino acids in renal proximal convoluted tubules of normal and streptozotocin-treated rats and rabbits.

A W Jans 1, R Willem 1
PMCID: PMC1133413  PMID: 2604695

Abstract

13C-n.m.r. spectroscopy was used to determine the metabolic fate of alanine and aspartate in rat and rabbit kidney proximal tubules. The main purpose of the present study was to investigate the effect of streptozotocin-induced diabetes on the influx of 13C label from [3-13C]alanine into the tricarboxylic acid cycle and through the fructose-1,6-bisphosphatase pathway. This influx was calculated from the relative enrichment of 13C in the various glutamate and glutamine carbon atoms. The relative proportion of 13C label which entered the tricarboxylic acid cycle via pyruvate carboxylase relative to the proportion that entered via pyruvate dehydrogenase was 1.92 +/- 0.02 in fed control rats and 2.27 +/- 0.04 in streptozotocin-treated rats. However, streptozotocin-induced diabetes did not significantly affect this ratio in rabbit proximal convoluted tubular cells. Only in rat proximal convoluted tubular cells did we observe an increase in flux through the fructose-1,6-bisphosphatase pathway by streptozotocin treatment compared with fed controls. The data suggest that streptozotocin-induced diabetes in rats causes the same metabolic changes as does chronic acidosis.

Full text

PDF
231

Selected References

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

  1. Balaban R. S., Soltoff S. P., Storey J. M., Mandel L. J. Improved renal cortical tubule suspension: spectrophotometric study of O2 delivery. Am J Physiol. 1980 Jan;238(1):F50–F59. doi: 10.1152/ajprenal.1980.238.1.F50. [DOI] [PubMed] [Google Scholar]
  2. Bárány M., Arús C., Chang Y. C. Natural-abundance 13C NMR of brain. Magn Reson Med. 1985 Jun;2(3):289–295. doi: 10.1002/mrm.1910020312. [DOI] [PubMed] [Google Scholar]
  3. Canioni P., Alger J. R., Shulman R. G. Natural abundance Carbon-13 nuclear magnetic resonance spectroscopy of liver and adipose tissue of the living rat. Biochemistry. 1983 Oct 11;22(21):4974–4980. doi: 10.1021/bi00290a015. [DOI] [PubMed] [Google Scholar]
  4. Cohen S. M. 13C NMR study of effects of fasting and diabetes on the metabolism of pyruvate in the tricarboxylic acid cycle and the utilization of pyruvate and ethanol in lipogenesis in perfused rat liver. Biochemistry. 1987 Jan 27;26(2):581–589. doi: 10.1021/bi00376a033. [DOI] [PubMed] [Google Scholar]
  5. Cohen S. M. Effects of insulin on perfused liver from streptozotocin-diabetic and untreated rats: 13C NMR assay of pyruvate kinase flux. Biochemistry. 1987 Jan 27;26(2):573–580. doi: 10.1021/bi00376a032. [DOI] [PubMed] [Google Scholar]
  6. Cohen S. M., Ogawa S., Shulman R. G. 13C NMR studies of gluconeogenesis in rat liver cells: utilization of labeled glycerol by cells from euthyroid and hyperthyroid rats. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1603–1609. doi: 10.1073/pnas.76.4.1603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cohen S. M., Rognstad R., Shulman R. G., Katz J. A comparison of 13C nuclear magnetic resonance and 14C tracer studies of hepatic metabolism. J Biol Chem. 1981 Apr 10;256(7):3428–3432. [PubMed] [Google Scholar]
  8. Cohen S. M. Simultaneous 13C and 31P NMR studies of perfused rat liver. Effects of insulin and glucagon and a 13C NMR assay of free Mg2+. J Biol Chem. 1983 Dec 10;258(23):14294–14308. [PubMed] [Google Scholar]
  9. FREEDMAN A. D., GRAFF S. The metabolism of pyruvate in the tricarboxylic acid cycle. J Biol Chem. 1958 Aug;233(2):292–295. [PubMed] [Google Scholar]
  10. Gstraunthaler G., Pfaller W., Kotanko P. Lack of fructose-1,6-bisphosphatase activity in LLC-PK1 cells. Am J Physiol. 1985 Jan;248(1 Pt 1):C181–C183. doi: 10.1152/ajpcell.1985.248.1.C181. [DOI] [PubMed] [Google Scholar]
  11. Guder W., Wiesner W., Stukowski B., Wieland O. Metabolism of isolated kidney tubules. Oxygen consumption, gluconeogenesis and the effect of cyclic nucleotides in tubules from starved rats. Hoppe Seylers Z Physiol Chem. 1971 Oct;352(10):1319–1328. doi: 10.1515/bchm2.1971.352.2.1319. [DOI] [PubMed] [Google Scholar]
  12. Gullans S. R., Brazy P. C., Dennis V. W., Mandel L. J. Interactions between gluconeogenesis and sodium transport in rabbit proximal tubule. Am J Physiol. 1984 Jun;246(6 Pt 2):F859–F869. doi: 10.1152/ajprenal.1984.246.6.F859. [DOI] [PubMed] [Google Scholar]
  13. Jans A. W., Kinne R. K. Characterization of the renal epithelial cell line, PKE 5, using 31P- and 13C-NMR spectroscopy. Biochim Biophys Acta. 1988 Apr 25;969(2):194–197. doi: 10.1016/0167-4889(88)90075-4. [DOI] [PubMed] [Google Scholar]
  14. Jans A. W., Leibfritz D. A 13C-NMR study on the influxes into the tricarboxylic acid cycle of a renal epithelial cell line, LLC-PK1/Cl4: the metabolism of [2-13C]glycine, L-[3-13C]alanine and L-[3-13C]aspartic acid in renal epithelial cells. Biochim Biophys Acta. 1988 Jul 29;970(3):241–250. doi: 10.1016/0167-4889(88)90123-1. [DOI] [PubMed] [Google Scholar]
  15. KOEPPE R. E., MOURKIDES G. A., HILL R. J. Some factors affecting routes of pyruvate metabolism in rats. J Biol Chem. 1959 Sep;234:2219–2222. [PubMed] [Google Scholar]
  16. Krebs H. A., Hems R., Weidemann M. J., Speake R. N. The fate of isotopic carbon in kidney cortex synthesizing glucose from lactate. Biochem J. 1966 Oct;101(1):242–249. doi: 10.1042/bj1010242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lancaster G., Mohyuddin F., Scriver C. R., Whelan D. T. A -aminobutyrate pathway in mammalian kidney cortex. Biochim Biophys Acta. 1973 Feb 28;297(2):229–240. doi: 10.1016/0304-4165(73)90069-x. [DOI] [PubMed] [Google Scholar]
  18. Nissim I., Yudkoff M., Segal S. Metabolic fate of glutamate carbon in rat renal tubules. Studies with 13C nuclear magnetic resonance and gas chromatography-mass spectrometry. Biochem J. 1987 Jan 15;241(2):361–370. doi: 10.1042/bj2410361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nissim I., Yudkoff M., Segal S. Metabolism of glutamine and glutamate by rat renal tubules. Study with 15N and gas chromatography-mass spectrometry. J Biol Chem. 1985 Nov 15;260(26):13955–13967. [PubMed] [Google Scholar]
  20. RICHTERICH R. W., GOLDSTEIN L. Distribution of glutamine metabolizing enzymes and production of urinary ammonia in the mammalian kidney. Am J Physiol. 1958 Nov;195(2):316–320. doi: 10.1152/ajplegacy.1958.195.2.316. [DOI] [PubMed] [Google Scholar]
  21. Wirthensohn G., Guder W. G. Renal substrate metabolism. Physiol Rev. 1986 Apr;66(2):469–497. doi: 10.1152/physrev.1986.66.2.469. [DOI] [PubMed] [Google Scholar]
  22. de los Santos C., Buldain G., Frydman B., Cannata J. J., Cazzulo J. J. Carbon-13 nuclear magnetic resonance analysis of [1-13C]glucose metabolism in Crithidia fasciculata. Evidence of CO2 fixation by phosphoenolpyruvate carboxykinase. Eur J Biochem. 1985 Jun 3;149(2):421–429. doi: 10.1111/j.1432-1033.1985.tb08942.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES