The nonenzymatic subunit of pseutarin C, a prothrombin activator from eastern brown snake (Pseudonaja textilis) venom, shows structural similarity to mammalian coagulation factor V.
Pseutarin C is a group C prothrombin activator from the venom of the eastern brown snake Pseudonaja textilis. It is a multi-subunit protein complex consisting of catalytic and nonenzymatic subunits similar to coagulation factor Xa and factor Va, respectively. Here we describe the complete sequence of the nonenzymatic subunit. Based on the partial amino acid sequence of the nonenzymatic subunit, degenerate primers were designed. Using a "walking" strategy based on sequentially designed primers, we determined the complete cDNA sequence of the nonenzymatic subunit. The cDNA encodes a protein of 1461 amino acid residues, which includes a 30-residue signal peptide, a mature protein of 1430 amino acid residues, and a stop codon. cDNA blot analysis showed a single transcript of approximately 4.6 kb. The deduced amino acid sequence shows approximately 50% identity to mammalian factor V and by homology has a similar domain structure consisting of domains A1-A2-B-A3-C1-C2. Interestingly, the B domain of pseutarin C is shorter than that of mammalian factor V (FV). Although most of the proteolytic activation sites are conserved, 2 of 3 proteolytic sites cleaved by activated protein C are mutated, and thus activated protein C is not able to inactivate this procoagulant toxin. The predicted posttranslational modifications, including disulfide bonds, N-glycosylation, phosphorylation, and sulfation, in pseutarin C are significantly different compared with bovine factor V. Thus, our data demonstrate that the nonenzymatic subunit of group C prothrombin activators is structurally similar to mammalian FV.
Department of Biological Sciences, Faculty of Science, National University of Singapore, Republic of Singapore.
Veena S
VS
Rao
Sanjay
S
Swarup
R Manjunatha
RM
Kini
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002417:Cattle; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000235:genetics; D004220:Disulfides / Q000378:metabolism; D004546:Elapid Venoms / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D005165:Factor V / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods; D051379:Mice; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D021122:Protein Subunits; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D004220:Disulfides; D004546:Elapid Venoms; D021122:Protein Subunits; C475049:Pseutarin C; D005165:Factor V