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Functional studies of yeast glucokinase

D Clifton1, R B Walsh, D G Fraenkel

  • 1Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115.

Journal of Bacteriology
|June 1, 1993
PubMed
Summary

In Saccharomyces cerevisiae, glucokinase activity does not increase glucose flux, even when hexokinases are absent. Glucose accumulates internally when external sources are depleted, suggesting a link between glucose entry and phosphorylation.

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Area of Science:

  • Biochemistry and Molecular Biology
  • Yeast Metabolism and Genetics

Background:

  • Glucose phosphorylation capacity typically exceeds glucose flux in wild-type Saccharomyces cerevisiae.
  • A mutant strain lacking hexokinases but retaining glucokinase presents a unique model for studying glucose metabolism.

Purpose of the Study:

  • To investigate the role of glucokinase in glucose transport and phosphorylation in Saccharomyces cerevisiae.
  • To determine if increased glucokinase activity can enhance glucose flux in the absence of hexokinases.

Main Methods:

  • Comparative analysis of glucose flux and internal glucose concentration in wild-type and glucokinase-dependent mutant strains.
  • Measurement of radioactive glucose uptake under various conditions, including glucose and maltose batch cultures.
  • Enzymatic activity assays of glucokinase and hexokinase following toluene treatment.

Main Results:

  • Internal glucose concentration remains low during active growth on glucose in both wild-type and mutant strains.
  • Additional glucokinase activity does not elevate glucose flux.
  • The glucokinase-dependent strain accumulates significant internal glucose after external glucose depletion or when grown on maltose.
  • Radioactive glucose uptake shows incomplete mixing with the internal pool in these accumulation scenarios.
  • Glucokinase activity is minimally detected after toluene treatment, unlike hexokinase.

Conclusions:

  • The findings suggest a potential link between glucose entry into the cell and its subsequent phosphorylation by glucokinase.
  • Separate mechanisms may explain the observed glucose accumulation and incomplete mixing phenomena.
  • Glucokinase's role in yeast glucose metabolism, particularly in relation to transport, warrants further investigation.

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