Amino acid sequence of trocarin, a prothrombin activator from Tropidechis carinatus venom: its structural similarity to coagulation factor Xa.
Among snake venom procoagulant proteins, group II prothrombin activators are functionally similar to blood coagulation factor Xa. We have purified and partially characterized the enzymatic properties of trocarin, the group II prothrombin activator from the venom of the Australian elapid, Tropidechis carinatus (rough-scaled snake). Prothrombin activation by trocarin is enhanced by Ca2+, phospholipids, and factor Va, similar to that by factor Xa. However, its amidolytic activity on peptide substrate S-2222 is significantly lower. We have determined the complete amino acid sequence of trocarin. It is a 46,515-Dalton glycoprotein highly homologous to factor Xa and shares the same domain architecture. The light chain possesses an N-terminal Gla domain containing 11 gamma-carboxyglutamic acid residues, followed by two epidermal growth factor (EGF)-like domains; the heavy chain is a serine proteinase. Both chains are likely glycosylated: the light chain at Ser 52 and the heavy chain at Asn 45. Unlike other types of venom procoagulants, trocarin is the first true structural homologue of a coagulation factor. It clots snake plasma and thus may be similar, if not identical, to snake blood coagulation factor Xa. Unlike blood factor Xa, it is expressed in high quantities and in a nonhepatic tissue, making snake venom the richest source of factor Xa-like proteins. It induces cyanosis and death in mice at 1 mg/kg body weight. Thus, trocarin acts as a toxin in venom and a similar, if not identical, protein plays a critical role in hemostasis.
Joseph
J S
JS
Bioscience Centre, Faculty of Science, the Department of Biochemistry, Faculty of Medicine, and the Bioprocessing Technology Centre, Faculty of Engineering, National University of Singapore, Singapore.
Chung
M C
MC
Jeyaseelan
K
K
Kini
R M
RM
D000595:Amino Acid Sequence; D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects; D002850:Chromatography, Gel; D002863:Chromogenic Compounds / Q000378:metabolism; D004546:Elapid Venoms / Q000201:enzymology*; D004789:Enzyme Activation / Q000187:drug effects; D015951:Factor Xa / Q000737:chemistry*; D006031:Glycosylation; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011516:Prothrombin / Q000737:chemistry* / Q000187:drug effects / Q000302:isolation & purification / Q000494:pharmacology / Q000633:toxicity; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
D002863:Chromogenic Compounds; D004546:Elapid Venoms; D009842:Oligopeptides; C120393:trocarin; C015608:benzoyl-Ile-Glu-Gly-Arg-p-nitroanilide; D011516:Prothrombin; D015951:Factor Xa