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

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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.

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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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Authors and Affiliations

  1. Institut für Botanik, Westfälische Wilhelms-Universität Münster, Schloßgarten 3, D-48149, Münster, Federal Republic of Germany

    K. B. Tenberge, C. Ruholl, M. Heinze & R. Eising

  2. Institut für Botanischer Garten, Westfälische Wilhelms-Universität, Münster

    K. B. Tenberge, C. Ruholl, M. Heinze & R. Eising

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  1. K. B. Tenberge
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  2. C. Ruholl
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  3. M. Heinze
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  4. R. Eising
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Tenberge, K.B., Ruholl, C., Heinze, M. et al. Purification and immuno-electron microscopical characterization of crystalline inclusions from plant peroxisomes. Protoplasma 196, 142–154 (1997). https://doi.org/10.1007/BF01279563

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  • DOI: https://doi.org/10.1007/BF01279563

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