This chapter describes preparation of geranylgeranylpyrophosphate synthetase from pig liver. Geranylgeranylpyrophosphate synthetase obtained from pig liver in a form free of farnesylpyrophosphate synthetase catalyzes the two reactions, and does not catalyze the condensation between dimethylallyl pyrophosphate and isopentenyl pyrophosphate. Assay methods and purification of geranylgeranylpyrophosphate synthetase are described in the chapter. The enzyme solution obtained by hydroxylapatite chromatography is concentrated by ultrafiltration, and chromatographed through a Sephadex G-200 column (3.0 × 80 cm) with 0.01 M phosphate buffer, pH 6.8. This chapter summarizes the results of purification up to hydroxylapatite chromatography starting with 200 g of pig liver.
ASJC Scopus subject areas
- Molecular Biology