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Differential Activation of Prothrombin by Factor Xa, Ecarin and Taipan Venom as Studied by ProteinChip Array Profiling with Surface Enhanced Laser Desorption Ionization (SELDI) Analysis
Author(s) -
Fareed Jawed,
Maddineni Jyothi,
Kujawski Michelle,
Jeske Walter,
Hoppensteadt Debra,
Bick Rodger
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.a1125-a
Subject(s) - chemistry , venom , snake venom , thrombin , prothrombin time , microbiology and biotechnology , biochemistry , immunology , biology , platelet , medicine
Bovine and human thrombins used for diagnostic testing and hemostatic purposes are typically made by activating prothrombin complexes with thromboplastins from varying sources. These products contain mainly α‐thrombin, though β‐ and γ‐thrombins are also formed. While potency of these products is standardized against reference material, no information on their molecular composition is provided. Recently, recombinant human thrombins have been developed by activating a precursor with factor Xa or snake venoms (ecarin and Taipan venom). It was hypothesized that cleavage of precursor products is dependent of the type of activator used. Purified human prothrombin or prothrombin complex concentrate was incubated with factor Xa, ecarin and Taipan venom. The cleavage products were profiled using SAX‐2 ProteinChip arrays employing SELDI detection (Ciphergen). Activation of purified prothrombin and prothrombin complexes resulted in unique biomarker profiles in the molecular weight range of 1–50 kDa. The proportion of specific peaks at 10–35 kDa differed. Moreover, the arrays differed significantly in the molecular weight range of < 10 kDa. Significant differences were apparent between the venom activated and factor Xa activated profiles. These studies suggest that the protease cleavage profile of prothrombin/prothrombin complexes results in characteristic cleavage products which may impact their biologic functions.