title: Group D prothrombin activators from snake venom are structural homologues of mammalian blood coagulation factor Xa.
abstract: Procoagulant venoms of several Australian elapids contain proteinases that specifically activate prothrombin; among these, Group D activators are functionally similar to coagulation factor Xa (FXa). Structural information on this class of prothrombin activators will contribute significantly towards understanding the mechanism of FXa-mediated prothrombin activation. Here we present the purification of Group D prothrombin activators from three Australian snake venoms (Hoplocephalus stephensi, Notechis scutatus scutatus and Notechis ater niger) using a single-step method, and their N-terminal sequences. The N-terminal sequence of the heavy chain of hopsarin D (H. stephensi) revealed that a fully conserved Cys-7 was substituted with a Ser residue. We therefore determined the complete amino acid sequence of hopsarin D. Hopsarin D shows approximately 70% similarity with FXa and approximately 98% similarity with trocarin D, a Group D prothrombin activator from Tropidechis carinatus. It possesses the characteristic Gla domain, two epidermal growth factor-like domains and a serine proteinase domain. All residues important for catalysis are conserved, as are most regions involved in interactions with factor Va and prothrombin. However, there are some structural differences. Unlike FXa, hopsarin D is glycosylated in both its chains: in light-chain residue 52 and heavy-chain residue 45. The glycosylation on the heavy chain is a large carbohydrate moiety adjacent to the active-site pocket. Overall, hopsarin D is structurally and functionally similar to mammalian coagulation FXa.
authors.affiliation: Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore 119260.
authors.forename: Veena S
authors.initials: VS
authors.lastname: Rao
authors.forename: Jeremiah S
authors.initials: JS
authors.lastname: Joseph
authors.forename: R Manjunatha
authors.initials: RM
authors.lastname: Kini
mesh_terms: D000581:Amidohydrolases / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D003029:Coagulants / Q000737:chemistry / Q000494:pharmacology; D004546:Elapid Venoms / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D015951:Factor Xa / Q000737:chemistry*; D018698:Glutamic Acid / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D006900:Hydroxylation; D008322:Mammals / Q000097:blood; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D011516:Prothrombin / Q000378:metabolism*; D020539:Sequence Analysis, Protein; D012697:Serine Endopeptidases / Q000737:chemistry / Q000378:metabolism; D012910:Snake Venoms / Q000737:chemistry*; D040681:Structural Homology, Protein
chemical_list: D003029:Coagulants; D004546:Elapid Venoms; D012910:Snake Venoms; C472031:notanarin D; C472032:notecarin D; D018698:Glutamic Acid; D011516:Prothrombin; D012697:Serine Endopeptidases; D015951:Factor Xa; D000581:Amidohydrolases; C472030:hopsarin D