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Journal of Biosciences

, Volume 9, Issue 1–2, pp 59–70 | Cite as

Uridine 5′-diphosphate glucose 4-epimerase from ehrlich ascites carcinoma cells

  • Samir Kumar Dutta
  • Manju Ray
  • Amar Bhaduri
Article

Abstract

Uridine 5′-diphosphate glucose 4-epimerase (EC 5.1.3.2) from Ehrlich ascites carcinoma cells was purified to apparent homogeneity using conventional procedures and NAD-hexane-agarose affinity chromatography. The protein had a molecular weight of 96,000. The ascites enzyme had an absolute requirement for exogenously added NAD (10 ΜM) for stability. This appears to be a unique feature of ascites epimerase since epimerase from other mammalian sources did not exhibit such a dependence. Exogenously added NAD was also needed for catalysis with an apparentK m value of 2.5 ΜM. NADH was a very potent competitive inhibitor (K i = 0.11 ΜM with respect to NAD) of the enzyme activity at pH values close to intracellular pH. The dependence of the enzyme on NAD for stability and its inhibition by NADH may have some potential significance in tumor metabolism

Keywords

Galactose metabolism UDP glucose 4-epimerase tumor 

Abbreviations used

UDP

Uridine 5′-diphosphate

buffer A

sodium phosphate buffer, pH 7.4, 20 mM, containing 2-mercaptoethanol l mM, EDTA 1 mM, NAD 10 ΜM

DEAE

diethylamino ethyl

EDTA

ethylene diamine tetra acetic acid (disodium salt)

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Copyright information

© Indian Academy of Sciences 1985

Authors and Affiliations

  • Samir Kumar Dutta
    • 1
  • Manju Ray
    • 1
  • Amar Bhaduri
    • 1
  1. 1.Division of Biochemistry, Department of PharmacyJadavpur UniversityCalcuttaIndia

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