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. 1984 Jul 15;221(2):303–309. doi: 10.1042/bj2210303

All hexokinase isoenzymes coexist in rat hepatocytes.

A Reyes, M L Cárdenas
PMCID: PMC1144040  PMID: 6089733

Abstract

The cellular distribution of hexokinase isoenzymes, N-acetylglucosamine Kinase and pyruvate kinases in rat liver was studied. Hepatocytes and non-parenchymal cells with high viability and almost no cross-contamination were obtained by perfusion in situ of the liver with collagenase, with the use of an enriched cell-culture medium in all steps of cell isolation. Separation of hexokinase isoenzymes was done by DEAE-cellulose chromatography, and enzyme activities were measured by a specific radioassay. Cytosol from isolated hepatocytes contained high-affinity hexokinases A, B and C, in addition to hexokinase D. The last-mentioned represented about 95% of total glucose-phosphorylating activity. Only hexokinase A was found associated t the particulate fraction. Isolated non-parenchymal cells contained only hexokinases A, B and C. N-Acetylglucosamine kinase was measured with a specific radioassay and was found as a single enzyme form in both hepatocytes and non-parenchymal cells, with higher activities in the former. Pyruvate kinase isoenzyme L was present only in the hepatocytes and isoenzyme K only in the non-parenchymal liver cells, confirming that they are good cellular markers.

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Selected References

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

  1. Allen M. B., Brockelbank J. L., Walker D. G. Apparent 'glucokinase' activity in non-hepatic tissues due to N-acetyl-D-glucosamine kinase. Biochim Biophys Acta. 1980 Aug 7;614(2):357–366. doi: 10.1016/0005-2744(80)90225-9. [DOI] [PubMed] [Google Scholar]
  2. Baur H., Kasperek S., Pfaff E. Criteria of viability of isolated liver cells. Hoppe Seylers Z Physiol Chem. 1975 Jun;356(6):827–838. doi: 10.1515/bchm2.1975.356.s1.827. [DOI] [PubMed] [Google Scholar]
  3. Berg T., Boman D. Distribution of lysosomal enzymes between parenchymal and Kupffer cells of rat liver. Biochim Biophys Acta. 1973 Oct 10;321(2):585–596. doi: 10.1016/0005-2744(73)90201-5. [DOI] [PubMed] [Google Scholar]
  4. Berry M. N., Friend D. S. High-yield preparation of isolated rat liver parenchymal cells: a biochemical and fine structural study. J Cell Biol. 1969 Dec;43(3):506–520. doi: 10.1083/jcb.43.3.506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bonney R. J., Walker P. R., Potter V. R. Isoenzyme patterns in parenchymal and non-parenchymal cells isolated from regenerating and regenerated rat liver. Biochem J. 1973 Dec;136(4):947–954. doi: 10.1042/bj1360947. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bontemps F., Hue L., Hers H. G. Phosphorylation of glucose in isolated rat hepatocytes. Sigmoidal kinetics explained by the activity of glucokinase alone. Biochem J. 1978 Aug 15;174(2):603–611. doi: 10.1042/bj1740603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  8. CRANE R. K., SOLS A. The association of hexokinase with particulate fractions of brain and other tissue homogenates. J Biol Chem. 1953 Jul;203(1):273–292. [PubMed] [Google Scholar]
  9. Craven P. A., Basford R. E. Brain hexokinase: immunohistochemical localization at the light microscopic level. Biochemistry. 1969 Sep;8(9):3520–3525. doi: 10.1021/bi00837a006. [DOI] [PubMed] [Google Scholar]
  10. Crisp D. M., Pogson C. I. Glycolytic and gluconeogenic enzyme activities in parenchymal and non-parenchymal cells from mouse liver. Biochem J. 1972 Feb;126(4):1009–1023. doi: 10.1042/bj1261009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cárdenas M. L., Rabajille E., Niemeyer H. Maintenance of the monomeric structure of glucokinase under reacting conditions. Arch Biochem Biophys. 1978 Sep;190(1):142–148. doi: 10.1016/0003-9861(78)90261-8. [DOI] [PubMed] [Google Scholar]
  12. Davagnino J., Ureta T. The identification of extrahepatic "glucokinase" as N-acetylglucosamine kinase. J Biol Chem. 1980 Apr 10;255(7):2633–2636. [PubMed] [Google Scholar]
  13. Dileepan K. N., Wagle S. R., Hofmann F., Decker K. Distribution profile of glucokinase and hexokinase in parenchymal and sinusoidal cells of rat liver during development. Life Sci. 1979 Jan 1;24(1):89–95. doi: 10.1016/0024-3205(79)90284-4. [DOI] [PubMed] [Google Scholar]
  14. EAGLE H. Amino acid metabolism in mammalian cell cultures. Science. 1959 Aug 21;130(3373):432–437. doi: 10.1126/science.130.3373.432. [DOI] [PubMed] [Google Scholar]
  15. Faulkner A., Jones C. T. The distribution of enzyme and isoenzyme activities between parenchymal and haematopoietic cells in the liver of the foetal guinea pig. Biochem J. 1979 Jan 15;178(1):89–95. doi: 10.1042/bj1780089. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Garnett M. E., Dyson R. D., Dost F. N. Pyruvate kinase isozyme changes in parenchymal cells of regenerating rat liver. J Biol Chem. 1974 Aug 25;249(16):5222–5226. [PubMed] [Google Scholar]
  17. González C., Ureta T., Sánchez R., Niemeyer H. Multiple molecular forms of ATP:hexose 6-phosphotransferase from rat liver. Biochem Biophys Res Commun. 1964 Jul 1;16(4):347–352. doi: 10.1016/0006-291x(64)90038-5. [DOI] [PubMed] [Google Scholar]
  18. Hernandez A., Crane R. K. Association of heart hexokinase with subcellular structure. Arch Biochem Biophys. 1966 Jan;113(1):223–229. doi: 10.1016/0003-9861(66)90176-7. [DOI] [PubMed] [Google Scholar]
  19. Hommes F. A., Everts R. S. Particulate and free hexokinase in fetal rat liver. Biol Neonate. 1978;33(3-4):193–200. doi: 10.1159/000241072. [DOI] [PubMed] [Google Scholar]
  20. Hue L., Bontemps F., Hers H. The effects of glucose and of potassium ions on the interconversion of the two forms of glycogen phosphorylase and of glycogen synthetase in isolated rat liver preparations. Biochem J. 1975 Oct;152(1):105–114. doi: 10.1042/bj1520105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jungermann K., Katz N. Functional hepatocellular heterogeneity. Hepatology. 1982 May-Jun;2(3):385–395. doi: 10.1002/hep.1840020316. [DOI] [PubMed] [Google Scholar]
  22. KATZEN H. M., SODERMAN D. D., NITOWSKY H. M. KINETIC AND ELECTROPHORETIC EVIDENCE FOR MULTIPLE FORMS OF GLUCOSE-ATP PHOSPHOTRANSFERASE ACTIVITY FROM HUMAN CELL CULTURES AND RAT LIVER. Biochem Biophys Res Commun. 1965 Apr 23;19:377–382. doi: 10.1016/0006-291x(65)90472-9. [DOI] [PubMed] [Google Scholar]
  23. Lagos R., Ureta T. Ontogeny of chick liver hexokinase isozymes. Arch Biol Med Exp (Santiago) 1981 Dec;14(4):331–341. [PubMed] [Google Scholar]
  24. MASTER R. W. POSSIBLE SYNTHESIS OF POLYRIBONUCLEOTIDES OF KNOWN BASE-TRIPLET SEQUENCES. Nature. 1965 Apr 3;206:93–93. doi: 10.1038/206093b0. [DOI] [PubMed] [Google Scholar]
  25. Rose I. A., Warms J. V. Mitochondrial hexokinase. Release, rebinding, and location. J Biol Chem. 1967 Apr 10;242(7):1635–1645. [PubMed] [Google Scholar]
  26. SIEKEVITZ P., POTTER V. R. Biochemical structure of mitochondria. II. Radioactive labeling of intra-mitochondrial nucleotides during oxidative phosphorylation. J Biol Chem. 1955 Jul;215(1):237–255. [PubMed] [Google Scholar]
  27. Sapag-Hagar M., Marco R., Sols A. Distribution of hexokinase and glucokinase between parenchymal and non-parenchymal cells of rat liver. FEBS Lett. 1969 Apr;3(1):68–71. doi: 10.1016/0014-5793(69)80099-2. [DOI] [PubMed] [Google Scholar]
  28. Seglen P. O. Preparation of isolated rat liver cells. Methods Cell Biol. 1976;13:29–83. doi: 10.1016/s0091-679x(08)61797-5. [DOI] [PubMed] [Google Scholar]
  29. Ureta T., Bravo R., Babul J. Rat liver hexokinases during development. Enzyme. 1975;20(6):334–348. doi: 10.1159/000458958. [DOI] [PubMed] [Google Scholar]
  30. Vera M. L., Cárdenas M. L., Niemeyer H. Kinetic, chromatographic and electrophoretic studies on glucose-phosphorylating enzymes of rat intestinal mucosa. Arch Biochem Biophys. 1984 Feb 15;229(1):237–245. doi: 10.1016/0003-9861(84)90149-8. [DOI] [PubMed] [Google Scholar]
  31. Werner H. V., Bartley J. C., Berry M. N. Glucose-adenosine 5'-triphosphate 6-phosphotransferases of isolated rat liver parenchymal cells. Biochem J. 1972 Dec;130(4):1153–1155. doi: 10.1042/bj1301153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. van Berkel T. J., Koster J. F., Hülsmann W. C. Distribution of L- and M-type pyruvate kinase between parenchymal and Kupffer cells of rat liver. Biochim Biophys Acta. 1972 Aug 28;276(2):425–429. doi: 10.1016/0005-2744(72)91003-0. [DOI] [PubMed] [Google Scholar]

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