Characterization and molecular cloning of two different type 2 ribosome-inactivating proteins from the monocotyledonous plant Polygonatum multiflorum.

Leaves of the monocotyledonous plant Polygonatum multiflorum L. (Solomon's seal) contain besides a monocot mannose-binding lectin two galactose/N-acetylgalactosamine (Gal/GalNAc)-binding type 2 ribosome-inactivating proteins (RIPs). Both RIPs were purified using a combination of classical protein purification techniques and affinity chromatography. Although both RIPs consist of protomers of 65 kDa, the P. multiflorum RIP monomer (PMRIPm) occurs as a monomer of approximately 60 kDa, whereas the tetramer (PMRIPt) is a tetramer of 240 kDa. Both RIPs exhibit similar RNA N-glycosidase activity but differ in their specific agglutination activity and carbohydrate-binding specificity, PMRIPt being a GalNAc-specific lectin whereas PMRIPm is Gal/GalNAc-specific. Toxicity tests indicated that both Polygonatum RIPs exhibit a very low cytotoxicity towards human and animal cells. Analysis of the genomic clones encoding both RIPs revealed a high degree of sequence similarity to other type 2 RIPs. Molecular modelling confirmed that both Polygonatum RIPs have a similar structure to ricin.