Isolation and partial characterization of a new ribosome-inactivating protein from Petrocoptis glaucifolia (Lag.) Boiss.

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.