Purification of Pyruvate Kinase from Germinating Castor Bean Endosperm

Overview

Journal Plant Physiol

Specialty Physiology

Date 1988 Apr 1

PMID 16666032

Citations 14

Authors

W C Plaxton

Affiliations

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Abstract

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.

Citing Articles

Transcriptional and Biochemical Characterization of Cytosolic Pyruvate Kinases in .

Wulfert S, Schilasky S, Krueger S Plants (Basel). 2020; 9(3).

PMID: 32168758 PMC: 7154858. DOI: 10.3390/plants9030353.

Pyruvate-kinase isoenzymes from zygotic and microspore-derived embryos of Brassica napus : Developmental profiles and subunit composition.

Sangwan R, Gauthier D, Turpin D, Pomeroy M, Plaxton W Planta. 2013; 187(2):198-202.

PMID: 24178043 DOI: 10.1007/BF00201938.

Leucoplast Pyruvate Kinase from Developing Castor Oil Seeds : Characterization of the Enzyme's Degradation by a Cysteine Endopeptidase.

Plaxton W Plant Physiol. 1991; 97(4):1334-8.

PMID: 16668552 PMC: 1081167. DOI: 10.1104/pp.97.4.1334.

Association of phosphoenolpyruvate phosphatase activity with the cytosolic pyruvate kinase of germinating mung beans.

Podesta F, Plaxton W Plant Physiol. 1991; 97(4):1329-33.

PMID: 16668551 PMC: 1081166. DOI: 10.1104/pp.97.4.1329.

Relationship between the Subunits of Leucoplast Pyruvate Kinase from Ricinus communis and a Comparison with the Enzyme from Other Sources.

Blakeley S, Plaxton W, Dennis D Plant Physiol. 1991; 96(4):1283-8.

PMID: 16668331 PMC: 1080927. DOI: 10.1104/pp.96.4.1283.

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