A NMR spectroscopic characterization of a new epithelial cell line, TALH-SVE, with properties of the renal medullary thick ascending limb of Henle's loop
Autor: | I. R. Von Recklinghausen, D. M. Scott, Arnold W. H. Jans |
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Jazyk: | angličtina |
Rok vydání: | 1991 |
Předmět: |
Uridine Diphosphate Glucose
Magnetic Resonance Spectroscopy Intracellular pH Citric Acid Cycle Biophysics Pyruvate Dehydrogenase Complex Biology Models Biological Epithelium Cell Line Phosphates chemistry.chemical_compound Animals Life Science Inositol Molecular Biology Pyruvate Carboxylase Alanine Kidney Medulla Perchlorates Cell Biology Nuclear magnetic resonance spectroscopy Metabolism Hydrogen-Ion Concentration Pyruvate dehydrogenase complex Biofysica Biochemistry chemistry Osmolyte Loop of Henle Rabbits Intracellular |
Zdroj: | Biochimica et biophysica acta-protein structure and molecular enzymology 1091 (1991) Biochimica et biophysica acta-protein structure and molecular enzymology, 1091, 179-187 |
ISSN: | 0167-4838 |
Popis: | NMR spectroscopy has been used to characterize a new renal cell line, TALH-SVE.1, which is derived from the medullary thick ascending limb of Henle's loop. From the 31P-NMR spectrum of a suspension of TALH-SVE cells using the chemical shift of the intracellular inorganic phosphate a value of 7.24 +/- 0.04 for the steady-state intracellular pH (pHi) was determined at pHo = 7.40. In addition, the 31P-NMR spectrum indicated rather high levels of UDPG, a finding confirmed by 1H-NMR spectra of perchloric acid extracts. The 1H-NMR data also demonstrate the presence of 'organic osmolytes' such as inositol, sorbitol, choline and glycerophosphoryl choline (GPC). 13C-NMR spectra of perchloric acid extracts of TALH-SVE cells incubated with [2-13C]- and [3-13 C]alanine were used to determine the relative influx in the Krebs cycle via pyruvate carboxylase (PCB) versus the influx via pyruvate dehydrogenase (PDH). The ratio was 0.41, while about 52% of all acetyl-CoA entering the Krebs cycle was unlabeled. 13C-NMR experiments also indicated that TALH-SVE cells lack gluconeogenic activity. The NMR study presented indicates that TALH-SVE cells possess metabolic pathways similar to those of the parental cells. |
Databáze: | OpenAIRE |
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