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  • Title: Broad-zone active-enzyme chromatography. Keto-acid dehydrogenases as associating systems.
    Author: Davis LC, Radke GA.
    Journal: Biophys Chem; 1983 Oct; 18(3):241-7. PubMed ID: 6688956.
    Abstract:
    We have extended the method of active-enzyme chromatography to include the use of broad zones of enzyme. This allows examination of interacting systems in a way formally analogous to sedimentation velocity so that simulation of the observed activity profiles is possible. The method has been applied using pyridine nucleotide-linked active enzyme assays. At the concentrations presently accessible by this technique, hexokinase and glucose-6-phosphate dehydrogenase, both associating systems, show single symmetrical boundaries, as does isolated diaphorase, while pyruvate and alpha-ketoglutarate dehydrogenases show more complex patterns, with the position of the reaction boundary for diaphorase activity being dependent on enzyme concentration.
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