Fragmentation of actin by thrombin-like snake venom proteases.

The effect of thrombin-like snake venom proteases (Ancrod of Agkistrodon rhodostoma and Batroxobins of Bothrops moojeni and Bothrops marajoensis) on skeletal muscle actin was studied and compared to the thrombic cleavage of this protein. Only EDTA-pretreated G- and F-actin were split by thrombin and Ancrod, while Batroxobins hydrolyzed native G-actin, too. The time course of digestion was followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. A split product of 37 500 daltons appeared first which was cleaved further resulting in three lower molecular weight fragments. The sodium dodecyl sulfate gel pattern of thrombic fragmentation was well distinguishable from those caused by Ancrod and Batroxobins. The first split products of Batroxobin digestion--a smaller peptide and the 37 500 dalton fragment--were isolated and by estimating their N-, and C-terminal end groups and amino acid compositions the peptide bond hydrolyzed first was located in the primary structure of actin. It was established that while thrombin split off two actino-peptides (at Arg(28)-Ala(29) and Arg(39)-His(40) from the N-terminal end of the molecule only Arg(39)-His(40) was cleaved by Batroxobins.