Preparation of Concanavalin A-Ricin A-Chain Conjugate and Its Biologic Activity Against Various Cultured Cells2

For the development of therapeutic agents that possess tissue-specific carriers, a method was devised to synthesize an artificial protein hybrid conjugate containing a moiety which binds to a cell membrane receptor and an active fragment of a toxic protein. By the introduction of an activated sulfhydryl group into concanavalin A (Con A), a conjugate of Con A and the ricin A-chain cross-linked with a disulfide linkage was synthesized. The purified conjugate was studied with regard to its inhibitory activity against protein synthesis in cell-free and cultured cell systems. The Con A-rich A-chain conjugate retained about one-third the inhibitory activity of ricin in a cell-free protein synthesis system. It also was highly toxic to cultured normal cells. These results indicate that the conjugate is a structural and functional analog of ricin and that the original membrane-binding chain (B-chain of ricin) could be replaced by Con A. Transformed cells were insensitive to this conjugate and required a longer preincubation time. The sensitivity of the normal cells was reduced in the presence of local anesthetics.