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Protoplasma

, Volume 196, Issue 3–4, pp 142–154 | Cite as

Purification and immuno-electron microscopical characterization of crystalline inclusions from plant peroxisomes

  • K. B. Tenberge
  • C. Ruholl
  • M. Heinze
  • R. Eising
Article

Summary

This paper describes the first purification method for crystalline inclusions (cores) from plant peroxisomes and an ultrastructural characterization of these isolated cores. 5-day-old sunflower (Helianthus annuus L.) cotyledon fractions which were highly enriched in cores showed negligible activity of the matrix enzyme glycolate oxidase but high catalase activity. As proven by electron microscopy, crystalline particles were surrounded neither by matrix material nor by membranes. Their geometrical outlines and ultrastructure were identical to those of cores in tissue sections, as was their reactivity with three different polyclonal catalase antibodies in the immunogold technique. Three-dimensional reconstruction, based on the geometrical outlines and ultrastructure of sectioned isolated cores from sunflower, suggested that they were quadrangular blocks. Ultrastructural analysis revealed an even periodic arrangement of repeating units which are probably cubes with 20 nm long edges. Isolated peroxisomal cores from potato (Solanum tuberosum L.) tubers had outlines which suggested that they were even rhomboidal prisms. They showed a granular ultrastructure without any repeating units and contained catalase, demonstrated by immunogold labelling and enzyme activity measurement. The results presented here suggested the hypothesis that the structural elements in plant peroxisomal cores are made of enzymatically active catalase, although the substructure may vary from species to species.

Keywords

Catalase Helianthus annuus Immunogold electron microscopy Peroxisomal cores Solanum tuberosum Ultrastructural analysis 

Abbreviations

ACOx

acyl-CoA oxidase

BSA

bovine serum albumin

EDTA

ethylenediamine-tetraacetate

GDH

glutamate dehydrogenase

GOx

glycolate oxidase

KPB

potassium phosphate buffer

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Copyright information

© Springer-Verlag 1997

Authors and Affiliations

  • K. B. Tenberge
    • 1
    • 2
  • C. Ruholl
    • 1
    • 2
  • M. Heinze
    • 1
    • 2
  • R. Eising
    • 1
    • 2
  1. 1.Institut für BotanikWestfälische Wilhelms-Universität MünsterMünsterFederal Republic of Germany
  2. 2.Institut für Botanischer GartenWestfälische Wilhelms-UniversitätMünster

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