Reference: Jacob L, et al. (1991) Purification and crystallization of yeast hexokinase isoenzymes. Characterization of different forms by chromatofocusing. J Chromatogr 587(1):85-92

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Abstract


The yeast hexokinase isoenzymes PI and PII have been purified in large amounts (20 mg) from overproducing yeast strains. The purification procedures of hexokinase PI and PII include anion-exchange chromatography on DEAE-Sephacel and chromatofocusing on PBE 94, hydrophobic interaction chromatography on phenyl-Sepharose (necessary for the isolation of the isoenzyme PI); in the final step either a Mono Q HR 5/5 or a Fractogel EMD TMAE 650(S) column was used. Hexokinase preparations were characterized before crystallization by chromatofocusing on a Mono P HR 5/20 FPLC column, where different forms of hexokinase can be rapidly distinguished by their elution behaviour. From both purified hexokinase PI and PII, large crystals were grown that diffract X-rays to high resolution.

Reference Type
Journal Article | Research Support, Non-U.S. Gov't
Authors
Jacob L, Beecken V, Bartunik LJ, Rose M, Bartunik HD
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