Journal
PHYTOCHEMISTRY
Volume 79, Issue -, Pages 27-38Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.phytochem.2012.04.001
Keywords
Phosphoglycerate kinase; Sunflower; Helianthus annuus; Seed; Oil; Fatty acids
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Funding
- Ministerio de Economia y Competitividad
- FEDER [AGL2011-23187]
- Advanta Seeds Company
- Natural Sciences and Engineering Research Council of Canada
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Three cDNAs encoding different phosphoglycerate kinase (PGK, EC 2.7.2.3) isoforms, two cytosolic (HacPCK1 and HacPCK2) and one plastidic (HapPGK), were cloned and characterized from developing sunflower (Helianthus annuus L.) seeds. The expression profiles of these genes showed differences in heterotrophic tissues, such as developing seeds and roots, where HacPGK1 was predominant, while HapPGK was highly expressed in photosynthetic tissues. The cDNAs were expressed in Escherichia colt, and the corresponding proteins purified to electrophoretic homogeneity, using immobilized metal ion affinity chromatography, and biochemically characterized. Despite the high level of identity between sequences, the HacPGK1 isoform showed strong differences in terms of specific activity, temperature stability and pH sensitivity in comparison to HacPGK2 and HapPGK. A polyclonal immune serum was raised against the purified HacPGK1 isoform, which showed cross-immunoreactivity with the other PGK isoforms. This serum allowed the localization of high expression levels of PGK isozymes in embryo tissues. (C) 2012 Elsevier Ltd. All rights reserved.
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