Characterization of three hemorrhagic factors from the venom of Okinawa habu (Trimeresurus flavoviridis)

Three hemorrhagic factors, HR1, HR2a and HR2b, of Okinawa habu venom were characterized in terms of their subunit structure, amino acid composition, metal content and immunological properties. HR1 is a dimer (mol. wt 90,000) consisting of two identical subunits at 25 degrees C, but polymerizes to form a tetramer at 4 degrees C. Two peaks corresponding to the dimer and the tetramer were observed upon ultracentrifugation analysis at 20 degrees C. HR2a and HR2b are monomers (mol. wt 24,000 and 19,000, respectively). HR1, HR2a and HR2b contain 407, 203 and 161 amino acids, respectively and the respective mol. wt based on the amino acid composition are 45,988, 23,075 and 18,457. The hemorrhagic factors contain Zn2+, Ca2+ and Mg2+, and were irreversibly inhibited by incubation with chelating reagents. The three hemorrhagic factors were immunologically distinguished from each other, and the hemorrhagic activities were inhibited by the respective antiserum. The activity of HR2a was also inhibited by the antiserum against HR2b.