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. 1990 Sep;94(1):291–296. doi: 10.1104/pp.94.1.291

Isolation of Chloroplastic Phosphoglycerate Kinase 1

Kinetics of the Two-Enzyme Phosphoglycerate Kinase/Glyceraldehyde-3-Phosphate Dehydrogenase Couple

Jerzy Macioszek 1,2,2, James B Anderson 1,2, Louise E Anderson 1,2
PMCID: PMC1077223  PMID: 16667700

Abstract

We report here a method for the isolation of high specific activity phosphoglycerate kinase (EC 2.7.2.3) from chloroplasts. The enzyme has been purified over 200-fold from pea (Pisum sativum L.) stromal extracts to apparent homogeneity with 23% recovery. Negative cooperativity is observed with the two enzyme phosphoglycerate kinase/glyceraldehyde-3-P dehydrogenase (EC 1.2.1.13) couple restored from the purified enzymes when NADPH is the reducing pyridine nucleotide, consistent with earlier results obtained with crude chloroplastic extracts (J Macioszek, LE Anderson [1987] Biochim Biophys Acta 892: 185-190). Michaelis Menten kinetics are observed when 3-phosphoglycerate is held constant and phosphoglycerate kinase is varied, which suggests that phosphoglycerate kinase-bound 1,3-bisphosphoglycerate may be the preferred substrate for glyceraldehyde-3-P dehydrogenase in the chloroplast.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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