Chemical modification of amino groups in cardiotoxin III from Taiwan cobra Naja naja atra) venom.

Cardiotoxin III (CTX III), a major cardiotoxin analogue isolated from the Taiwan cobra (Naja naja atra) venom was modified, either with trinitrobenzene sulfonate (TNBS) or 4-chloro-3,5-dinitrobenzoate (CDNB). Under the conditions of limited reagent availability, three mono-TNP derivatives modified at Lys-5, 12, or 44, and three mono-CDNP derivatives at Lys-12, 23, or 44 were isolated, respectively. The biological activities of CTX III were more or less affected after each of these reactive amino groups were modified. In particular, the hemolytic activity to human erythrocytes and cytotoxicity on NS-1 cells of CTX III decreased to 31% and 50%, respectively, when Lys-12 was trinitrophenylated. More pronounced alteration in these activities was observed as this amino group was carboxydinitrophenylated. A good correlation between the hemolytic activity and cytotoxicity was found. These results indicate that epsilon-amino group at Lys-12 is most closely related to the hemolytic and cytotoxic activities of CTX III. The antigenicity of modified derivatives still remained intact as measured by ELISA.