Purification of Antiviral Proteins with Ribosome-Inactivating Properties from Plants

Max Schönfelder, Ute Janott, Roland Frötschl, Karl-Wolfgang Mundry, Günter Adam
1992 Zeitschrift für Naturforschung C - A Journal of Biosciences  
A generally applicable highly effective purification method for antiviral plant proteins, selective for those with ribosome-inactivating properties, was developed. It involved affinity chromatography on Cibacron Blue Sepharose of the extracted plant sap after acid-ethanol precipitation and finally cation exchange chromatography on Mono S (Pharmacia). After the second chromatography step electrophoretically pure proteins from the four plant species, Chenopodium amaranticolor, Dianthus barbatus,
more » ... Dianthus barbatus, Phytolacca americana and Spinacia oleracea, were obtained. These proteins inhibited infection of test plants with alfalfa mosaic virus and in vitro translation of TMV-RNA. The apparent molecular weights of the purified proteins were determined by SDS polyacrylamide gel electrophoresis as: 29 kDa for C. amaranticolor, 33 kDa for D. barbatus, 29 kDa for P. americana and 36 kDa for S. oleracea. Comparisons of the proteins by indirect ELISA and by Western blotting with polyclonal antisera from rabbits revealed that all four proteins were serologically related but not identical. In addition all four proteins were found to be glycosylated.
doi:10.1515/znc-1992-9-1016 fatcat:6pdk5sixnjgmbbjdircbvvr7ea