Purification and characterization of ostrich prothrombin

Carminita Frost, Ryno Naudé, Willem Oelofsen, Koji Muramoto, Takako Naganuma, Tomohisa Ogawa

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)

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 Cu2+-chelate Sepharose chromatography. Ostrich prothrombin exhibited a Mr 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 β-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.

Original languageEnglish
Pages (from-to)1151-1159
Number of pages9
JournalInternational Journal of Biochemistry and Cell Biology
Volume32
Issue number11-12
DOIs
Publication statusPublished - 2000

Keywords

  • Blood coagulation
  • Molecular evolution
  • Ostrich
  • Prothrombin
  • cDNA

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

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