title: Purification and characterization of ostrich prothrombin.
abstract: The work focused on the penultimate enzyme, prothrombin, in the coagulation cascade. Prothrombin was purified and characterized from ostrich plasma. The results obtained contribute to a better understanding of blood coagulation in the ostrich and the evolution of prothrombin and the coagulation cascade. Prothrombin was purified from ostrich plasma by barium chloride precipitation, ammonium sulfate fractionation, and DEAE-cellulose and Cu(2+)-chelate Sepharose chromatography. Ostrich prothrombin exhibited a M(r) of 72,800 and a pI of 6.9 using SDS-PAGE and PAG-isoelectrofocusing, respectively. The N-terminal sequence of ostrich prothrombin showed 78 and 87% identity with human and bovine, respectively. The cDNA was isolated from ostrich liver and the predicted amino acid sequence compared with those from other species. Ostrich prothrombin shares sequence identity with chicken (84%), human (60%), bovine (59%), rat (60%), mouse (59%) and hagfish (50%) prothrombin, suggesting a common function of prothrombin in these vertebrates. Amino acid sequence identities indicate that the thrombin beta-chain (62%) and the propeptide-Gla (75%) domains are the regions most constrained for the common functions of vertebrate prothrombins. Ostrich prothrombin, therefore, shows similarity in structure to other vertebrate prothrombins.
authors.affiliation: Department of Biochemistry and Microbiology, University of Port Elizabeth, PO Box 1600, Port Elizabeth 6000, South Africa.
authors.forename: C
authors.initials: C
authors.lastname: Frost
authors.forename: R
authors.initials: R
authors.lastname: Naudé
authors.forename: W
authors.initials: W
authors.lastname: Oelofsen
authors.forename: K
authors.initials: K
authors.lastname: Muramoto
authors.forename: T
authors.initials: T
authors.lastname: Naganuma
authors.forename: T
authors.initials: T
authors.lastname: Ogawa
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001777:Blood Coagulation; D019143:Evolution, Molecular; D006801:Humans; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011516:Prothrombin / Q000737:chemistry* / Q000302:isolation & purification*; D016415:Sequence Alignment; D020310:Struthioniformes / Q000097:blood*
chemical_list: D011516:Prothrombin