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. 1983 Jun 1;211(3):617–623. doi: 10.1042/bj2110617

Purification, properties and assay of D-ribulose 5-phosphate 3-epimerase from human erythrocytes.

A Karmali, A F Drake, N Spencer
PMCID: PMC1154406  PMID: 6882362

Abstract

A direct assay procedure is described for D-ribulose 5-phosphate 3-epimerase (EC 5.1.3.1) which exploits differences in the c.d. spectra of substrate and product. The enzyme has been purified from human erythrocytes and was resolved by gel filtration and sucrose-density-gradient centrifugation into a major component of apparent Mr 45 000 and a minor component of Mr 23 000. Electrophoresis in sodium dodecyl sulphate gave a single component corresponding to Mr 23 000. Kinetic and sucrose-density-gradient centrifugation data indicate dissociation of the dimeric form of the enzyme into monomers of low specific activity; substrate favours the active dimeric form of the enzyme. At concentrations of the enzyme where both forms of the enzyme are present initial velocity data yielded a Hill plot with an interaction coefficient of approx. 2.0, indicating co-operative binding of substrate under these conditions.

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Selected References

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