PMID: 11322889Apr 27, 2001Paper

Purification and cloning of chloroplast 6-phosphogluconate dehydrogenase from spinach. Cyanobacterial genes for chloroplast and cytosolic isoenzymes encoded in eukaryotic chromosomes

European Journal of Biochemistry
K KrepinskyC Schnarrenberger

Abstract

Previous attempts to purify chloroplast 6-phosphogluconate dehydrogenase (cp6PGDH), a key enzyme of the oxidative pentose phosphate pathway, have been unsuccessful due to rapid activity loss. An efficient purification protocol was developed and the enzyme from spinach leaves was purified 1000-fold to apparent homogeneity with a specific activity of 60 U.mg-1. The enzyme is a homodimer with subunits of 50 kDa. Antibodies raised against the purified cp6PGDH detected a 53-kDa protein from a crude extract, indicating alterations during purification. Purified cp6PGDH was microsequenced and the corresponding spinach cDNA was cloned using PCR techniques and degenerate primers. The cDNA for cytosolic 6PGDH from spinach was cloned for comparison. Phylogenetic analysis in the context of available homologues from eukaryotes and eubacteria revealed that animal and fungal cytosolic 6PGDH sequences are more similar to their homologues from gamma-proteobacteria, whereas plant 6PGDH is more similar to its cyanobacterial homologues. The ancestral gene for higher plant 6PGDH was acquired from the antecedent of plastids through endosymbiosis and gene transfer to the nucleus. A subsequent gene duplication gave rise to higher plant cytosolic 6PGDH,...Continue Reading

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Citations

Dec 31, 2003·Molecular and Biochemical Parasitology·Mariana Igoillo Esteve, Juan José Cazzulo
Jun 12, 2002·Biochemical and Biophysical Research Communications·José R Pérez-CastiñeiraAurelio Serrano
Dec 7, 2002·Molecular and Biochemical Parasitology·Charles L GreenblattMichael P Barrett
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Mar 27, 2013·Journal of Experimental Botany·Gertraud SpielbauerA Mark Settles
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Jan 2, 2003·Proceedings of the National Academy of Sciences of the United States of America·Yves BalmerBob B Buchanan
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Jan 29, 2003·Proceedings of the National Academy of Sciences of the United States of America·William Martin, Piet Borst
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Jul 2, 2016·Plant Signaling & Behavior·Marie-Christin Lutterbey, Antje von Schaewen
May 28, 2016·Scientific Reports·Joana FariaAnabela Cordeiro-da-Silva
May 7, 2013·International Journal of Molecular Sciences·Rosa M MateosJosé M Palma

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