Resolution of subunits of thrombin like enzyme Agapurin from Agkistrodon halys venom and sequencing of amino acid residues

[Abstract] Objective To determine the N-terminal 15 amino acid residues of two similar subunits of Agacutin. The two subunits of the enzyme molecule need to be split to obtain the subunit single peptide chain and sequenced. Methods Highly purified Agacutin was obtained by biochemical method. Single subunit samples for identification and sequencing were prepared by SDS-PAGE electrophoresis separation, subunit adhesive strip recovery and PVDF membrane electrotransfer technology. The latter determined 15 amino acid residues at the N-terminal of subunit by Edman degradation method. Results The sequence of large subunit (16 kDa) was DCSSGWSSYEEHQYY, and that of small subunit (15 kDa) was DSSGWSSYEGHEYYV. Conclusion The sequencing results showed that Agacutin was a new snake venom thrombin like enzyme by searching NCBI database.