Characterization of a potent platelet aggregation inducer from Cerastes cerastes (Egyptian sand viper) venom.
A potent, proteinaceous inducer of platelet aggregation designated as IVa, has been purified to homogeneity from Cerastes cerastes venom by molecular sieve and ion exchange chromatography. It is composed of 2 subunits with total M(r) of 62,000 as shown by native gel chromatography and chemical cross-linking with disuccinimidyl suberate. It is not clear at the present time whether both subunits are identical gene products, however, both have identical N-terminal sequences for the first 15 amino acids. The protein has a pI above 9.6. IVa (0.1 micrograms/ml) could aggregate platelets up to 80% and was inhibited by p-APMSF, leupeptin, iodoacetamide, protein kinase C inhibitor, phosphatase inhibitor, ATP and PGE1, while it was insensitive to acetylsalicylic acid, ADP scavenger system, protein kinase A inhibitor and hirudin. Protein IVa is a serine proteinase with thrombin-like activity as it hydrolysed thrombin chromogenic substrate CBS 34.47, its aggregatory activity was partially inhibited by monoclonal antibodies against GPIb and the thrombin receptor, as was the thrombin, and its ability to induce intracellular Ca2+ release was blocked by pretreating platelets with thrombin. Unlike thrombin, the IVa protein showed very weak coagulant activity as indicated by plasma recalcification time and fibrinogen clotting time although it could hydrolyse fibrinogen alpha-chains.
Basheer
A R
AR
Department of Biological Chemistry, Medical College of Pennsylvania, Hahnemann University, Philadelphia 19102-1192, USA.
el-Asmar
M F
MF
Soslau
G
G
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001777:Blood Coagulation / Q000187:drug effects; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010974:Platelet Aggregation / Q000187:drug effects*; D011489:Protein Denaturation; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D014757:Viper Venoms / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000494:pharmacology
D000596:Amino Acids; D014757:Viper Venoms; D012697:Serine Endopeptidases