Purification of Pyruvate Kinase from Germinating Castor Bean Endosperm 1
AUTOR(ES)
Plaxton, William C.
RESUMO
Cytosolic pyruvate kinase from endosperm of germinating castor beans (Ricinus communis L.; cv Hale) has been purified 3100-fold to apparent homogeneity and a final specific activity of 203 micromole pyruvate produced/minute per milligram protein. Purification steps included: heat treatment, polyethylene glycol fractionation, Q-Sepharose, ADP-agarose, Mono-Q and Phenyl Superose chromatography. Nondenaturing polyacrylamide gel electrophoresis of the final sample resulted in a single protein staining band which co-migrated with pyruvate kinase activity. Two protein staining bands of 57 and 56 kilodaltons were observed following SDS polyacrylamide gel electrophoresis of the final preparation. The native molecular mass was found to be about 240 kilodaltons. This enzyme appears to be a tetramer composed of two different subunits. The presence of dithioerythritol (2 millimolar) was required for optimal activity of the purified enzyme.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1054628Documentos Relacionados
- Pyruvate Dehydrogenase Complex from Germinating Castor Bean Endosperm 1
- Characterization and Kinetics of Isoenzymes of Pyruvate Kinase from Developing Castor Bean Endosperm 1
- Lipid Composition of Organelles from Germinating Castor Bean Endosperm 1
- Membrane Lipid Metabolism in Germinating Castor Bean Endosperm 1
- ATP Citrate Lyase from Germinating Castor Bean Endosperm: Localization and some Properties 1