A thrombin inhibitor from the gut of Boophilus microplus ticks.
A thrombin inhibitor was identified for the first time in the gut of the cattle tick Boophilus microplus. Here we present the partial purification and characterization of this new molecule, which was purified from the gut extract by three chromatographic steps: ion-exchange, gel filtration and affinity chromatography in a thrombin-Sepharose resin. In SDS-PAGE the inhibitor showed an apparent molecular mass of circa 26 kDa, which is different from the two thrombin inhibitors present in the saliva of this tick. The new inhibitor delays bovine plasma clotting time and inhibits both thrombin induced fibrinogen clotting and thrombin induced platelet aggregation. However, it does not interfere with thrombin amidolytic activity upon a small substrate (H-D-Phe-Pip-Arg-para-nitroanilide), which does not require binding to thrombin exosites. Therefore, the inhibitor does not block thrombin active site, although it must interfere with one of the thrombin exosites. B. microplus gut thrombin inhibitor (BmGTI) is also capable of enhancing activated protein C (APC) activity upon its specific substrate (H-D-Glu-Pro-Arg-para-nitroanilide), an activity never described before among B. microplus molecules.
Ricci
Clarisse Gravina
CG
Centro de Biotecnologia da Universidade Federal do Rio Grande do Sul, 15005, Porto Alegre, 91501-970, Brazil.
Pinto
Antônio Frederico Michel
AF
Berger
Markus
M
Termignoni
Carlos
C
D000818:Animals; D000925:Anticoagulants / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D041981:Gastrointestinal Tract / Q000737:chemistry; D048494:Rhipicephalus / Q000737:chemistry*; D015842:Serine Proteinase Inhibitors / Q000032:analysis; D013917:Thrombin / Q000037:antagonists & inhibitors*
D000925:Anticoagulants; D015842:Serine Proteinase Inhibitors; D013917:Thrombin
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