Abstract
Petrocoptis glaucifolia, a paleoendemic member of the Caryophyllaceae from the North of Spain, was found to contain at least five proteins that inhibit protein synthesis in a rabbit reticulocyte lysate. One of them, for which the name petroglaucin is proposed, was purified to apparent electrophoretic homogeneity by chromatography through S-Sepharose Fast Flow, Sephadex G-75 and CM-Sepharose Fast Flow. The apparent Mr of the preparation was 27500. This protein does not contain appreciable glycan chains and displays 45.8% of NH2-terminal amino-acid sequence homology with some ribosome-inactivating proteins from Saponaria officinalis, another member of the Caryophyllaceae. Petroglaucin shows the following functional properties: (i) it strongly inhibits the rabbit-reticulocyte-lysate system and Vicia sativa cell-free extracts, both coded by endogenous messengers, and also inhibits poly(U)-directed polyphenylalanine synthesis by Vicia sativa cell-free extracts and purified rat-liver ribosomes; (ii) it shows much less inhibitory capacity in wheat-germ, Cucumis sativus and rat-liver cell-free systems coded by endogenous messengers; (iii) the inhibitory effects on purified rat-liver ribosomes were irreversible; (vi) it promotes the release of adenine from purified rat-liver ribosomes. The total activity of this translational inhibitor has been found to increase up to 11-fold during its purification, indicating that some regulatory factor that normally blocks the translational inhibitory activity of the ribosome-inactivating protein in crude extracts of the plant is removed during purification.
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Abbreviations
- DFA:
-
dimethylforrnamide
- PAGE:
-
polyacrylamide gel electrophoresis
- poly(U):
-
polyuridylic acid
- RIP:
-
ribosome inactivating protein
- SDS:
-
sodium dodecyl sulfate
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The work in Valladolid was supported by grants from CICYT (BIO 88-0705), Junta de Castilla y León and Iberduero S.A. The work in Bologna was supported by grants from Ministerio della Pubblice Istruzione, Associazione Italiana per la Ricerca sul Cancro, Consiglio Nazionale dell Ricerche, within the Progetto finalizzato Biotenologia e biostrumentazione. F.J. Arias and M.A. Rojo hold fellowships from Iberduero S.A. J.M. Ferreras and R. Iglesias hold postdoctoral fellowships from Ministerio de Educación y Ciencia. We thank Professor R. Parilla (Instituto Gregorio Marañon, Madrid) for his critical reading of the manuscript. The supervision of the English version of the manuscript by N. Skinner is greatly appreciated.
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Arias, F.J., Rojo, M.A., Ferreras, J.M. et al. Isolation and partial characterization of a new ribosome-inactivating protein from Petrocoptis glaucifolia (Lag.) Boiss. Planta 186, 532–540 (1992). https://doi.org/10.1007/BF00198033
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DOI: https://doi.org/10.1007/BF00198033