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Wheat ribosome-inactivating proteins: Seed and leaf forms with different specificities and cofactor requirements

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Abstract

Distinct forms of ribosome-inactivating proteins were purified from wheat (Triticum aestivum L.) germ and leaves and termed tritin-S and tritin-L, respectively. These differ in size and charge and are antigenically unrelated. They are both RNA N-glycosidases which act on 26S rRNA in native yeast (Saccharomyces cerevisiae) ribosomes by the removal of A3024 located in a universally conserved sequence in domain VII which has previously been identified as the site of action of ricin A-chain. Tritin-S and tritin-L differ in both their ribosome substrate specificities and cofactor requirements. Tritin-S shows only barely detectable activity on ribosomes from the endosperm, its tissue of synthesis, whereas tritin-L is highly active on leaf ribosomes. Additionally, tritin-S is inactive on wheat germ, tobacco leaf and Escherichia coli ribosomes but active on rabbit reticulocyte and yeast ribosomes. Tritin-L is active on ribosomes from all of the above sources. Tritin-S, unlike tritin-L shows a marked requirement for ATP in its action.

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Abbreviations

CM:

carboxymethyl

FPLC:

fast protein liquid chromatography

NEPHGE:

non-equilibrium pH gradient gel electrophoresis

PAP:

pokeweed antiviral protein

RIP:

ribosome-inactivating protein

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Authors and Affiliations

  1. Department of Biological Sciences, University of Warwick, CV4 7AL, Coventry, West Midlands, UK

    Andrea J. Massiah & Martin R. Hartley

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  1. Andrea J. Massiah
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  2. Martin R. Hartley
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A.J.M. was the recipient of a U.K. Science and Engineering Research Council CASE studentship sponsored by Agricultural Genetics Company Ltd., Cambridge CB4 4GG, UK.

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Massiah, A.J., Hartley, M.R. Wheat ribosome-inactivating proteins: Seed and leaf forms with different specificities and cofactor requirements. Planta 197, 633–640 (1995). https://doi.org/10.1007/BF00191571

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  • DOI: https://doi.org/10.1007/BF00191571

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