Determination of the Deoxyglucose and Glucose Phosphorylation Ratio and the Lumped Constant in Rat Brain and a Transplantable Rat Glioma

Rajesh Kapoor, Alexander M. Spence, Mark Muzi, Michael M. Graham, Gregory L. Abbott, Kenneth A. Krohn

Research output: Contribution to journalArticlepeer-review

37 Scopus citations

Abstract

Abstract: Mitochondrially bound hexokinase (ATP‐D‐hexose‐6‐phosphotransferase; EC 2.7.1.1) was dissociatively extracted from normal rat brains and intracerebral and subcutaneous implants of the 36B‐10 glioma. At least 70% of the total hexokinase enzyme activity in normal and glioma tissue was associated with the mitochondrial fraction. Purification of the crude tissue extracts by ion‐exchange and affinity chromatography followed by analysis with sodium dodecyl sul‐fate‐polyacrylamide gel electrophoresis revealed a successive purification of the enzyme to homogeneity with a molecular size of 98 kilodaltons. Enzyme kinetics with glucose or 2‐deoxyglucose (2‐DG) as the substrate were measured spec‐trophotometrically by coupling the appropriate reactions to either NADPH or NAD+ formation. The Km of hexokinase with glucose as the substrate in the intracerebral glioma (0.138 m.M) and subcutaneous glioma (0.183 mM) tissues was 2.1 ‐2.7‐fold higher than that observed in normal brain tissue (0.067 mM) (p < 0.001). No significant differences were observed in the Km for hexokinase with 2‐DG as the substrate in the glioma and normal brain tissue. The phosphorylation ratio for normal brain was 0.320 and was increased in the intracerebral glioma to 0.694 and in the subcutaneous glioma to 0.519. The ratios of deoxyglucose and glucose volumes of distribution in normal brain and intracerebral glioma tissues were 1.70 and 1.85, respectively. The lumped constants calculated directly from the phosphorylation ratios and the volumes of distribution of deoxyglucose and glucose were 0.517 in normal brain and 1.168 in intracerebral glioma. Our results indicate the lumped constant is increased 2.26‐fold in intracerebral glioma compared with normal brain.

Original languageEnglish (US)
Pages (from-to)37-44
Number of pages8
JournalJournal of neurochemistry
Volume53
Issue number1
DOIs
StatePublished - Jul 1989

Keywords

  • 2‐Deoxyglucose
  • Enzyme kinetics
  • Glioma
  • Glucose
  • Hexokinase
  • Lumped constant

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

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