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McMorrow EM, Bradbeer JW. Separation, purification, and comparative properties of chloroplast and cytoplasmic phosphoglycerate kinase from barley leaves. Plant Physiol. 1990;93(2):374-83doi: 10.1104/pp.93.2.374.
McMorrow, E. M., & Bradbeer, J. W. (1990). Separation, purification, and comparative properties of chloroplast and cytoplasmic phosphoglycerate kinase from barley leaves. Plant physiology, 93(2), 374-83. https://doi.org/10.1104/pp.93.2.374
McMorrow, E M, and Bradbeer, J W. "Separation, purification, and comparative properties of chloroplast and cytoplasmic phosphoglycerate kinase from barley leaves." Plant physiology vol. 93,2 (1990): 374-83. doi: https://doi.org/10.1104/pp.93.2.374
McMorrow EM, Bradbeer JW. Separation, purification, and comparative properties of chloroplast and cytoplasmic phosphoglycerate kinase from barley leaves. Plant Physiol. 1990 Jun;93(2):374-83. doi: 10.1104/pp.93.2.374. PMID: 16667476; PMCID: PMC1062521.
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Plant Physiol. 1990 Jun;93(2):374-83. doi: 10.1104/pp.93.2.374.

Separation, purification, and comparative properties of chloroplast and cytoplasmic phosphoglycerate kinase from barley leaves.

Plant physiology

E M McMorrow, J W Bradbeer

Affiliations

  1. Division of Biosphere Sciences, King's College London, Campden Hill Road, London W8 7AH, United Kingdom.

PMID: 16667476 PMCID: PMC1062521 DOI: 10.1104/pp.93.2.374

Abstract

The chloroplast and cytoplasmic isoenzymes of phosphoglycerate kinase (PGK) (EC. 2.7.2.3) from Hordeum vulgare leaves have been separated and purified for the first time to apparent homogeneity. The method for purifying the isoenzymes is described here and consists of DEAE Sephacel chromatography followed by affinity chromatography on ATP Sepharose. This consistently provided a 500- to 900-fold purification of each isoenzyme. Most of the total PGK in green barley leaves was found to be in the chloroplasts with only 10% in the cytoplasm. The immunological properties of the two isoenzymes were compared. The antisera raised to the separate isoenzymes showed cross-reactivity, although there is evidence that each isoenzyme possesses some distinct epitopes. The isoenzymes differ in overall charge with isoelectric points at 5.2 and 5.4 for the chloroplast and cytoplasmic isoenzymes, respectively. Molecular mass estimations by gel filtration and sodium dodecyl sulfate polyacrylamide gel electrophoresis provided similar values of approximately 38 kilodaltons for each isoenzyme, some 4 to 5 kilodaltons less than the values calculated from the cDNA sequences of the wheat isoenzymes. The isoenzymes have broadly similar pH optima of pH 7 to 8. The cytoplasmic isoenzyme is more thermally stable than the chloroplast isoenzyme. Further studies are now in progress to compare both the regulatory properties of the isoenzymes and also their three-dimensional structures as compared with the yeast enzyme.

References

  1. J Biol Chem. 1953 Oct;204(2):939-48 - PubMed
  2. Biochim Biophys Acta. 1975 Mar 28;386(1):181-95 - PubMed
  3. J Biol Chem. 1987 Oct 25;262(30):14472-8 - PubMed
  4. Eur J Biochem. 1983 Oct 17;136(1):101-6 - PubMed
  5. Eur J Biochem. 1978 Apr 17;85(2):493-501 - PubMed
  6. Plant Physiol. 1975 Feb;55(2):168-71 - PubMed
  7. Anal Biochem. 1982 Dec;127(2):316-21 - PubMed
  8. Nucleic Acids Res. 1989 Aug 25;17(16):6569-80 - PubMed
  9. Biochim Biophys Acta. 1976 Aug 12;445(1):74-88 - PubMed
  10. Plant Physiol. 1982 Apr;69(4):825-8 - PubMed
  11. Plant Physiol. 1986 Feb;80(2):301-4 - PubMed
  12. Eur J Biochem. 1976 Apr 1;63(2):483-90 - PubMed
  13. Plant Physiol. 1970 May;45(5):583-5 - PubMed
  14. Scand J Immunol Suppl. 1973;1:101-3 - PubMed
  15. Biochim Biophys Acta. 1967 Jan 11;132(1):33-40 - PubMed
  16. Biochem Biophys Res Commun. 1979 May 28;88(2):515-21 - PubMed
  17. EMBO J. 1982;1(12):1635-40 - PubMed
  18. Eur J Biochem. 1978 Apr 17;85(2):503-16 - PubMed

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