Notecarin D Binds Human Factor V and Factor Va with High Affinity in the Absence of Membranes
作者： Jennifer L. Newell-Caito , Malabika Laha , Anthony C. Tharp , Jonathan I. Creamer , Hong Xu
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摘要: Notecarin D (NotD) is a prothrombin (ProT) activator in the venom of tiger snake, Notechis scutatus, and factor Xa (FXa) homolog. NotD binds specifically to FXa binding site expressed on V (FV) upon activation Va (FVa) by thrombin. active site-labeled with 5-fluorescein ([5F]FFR-NotD) FV FVa remarkably high affinity absence phospholipids (KD 12 ≤ 0.01 nm, respectively). In presence membranes, [5F]FFR-NotD for similar, but increased ∼55-fold FV. Binding Oregon Green ∼5,000- ∼80-fold weaker than [5F]FFR-NotD, respectively. reports not 3-fold increase tripeptide substrate hydrolysis, demonstrating allosteric regulation FVa. The NotD·FVa·membrane complex activates ProT Km(app) similar prothrombinase, ∼85-fold without membranes. Active site-blocked exhibits potent anticoagulant activity plasma thrombin generation assays, representing inhibition productive prothrombinase assembly possible disruption tissue pathway inhibitor. results show that membrane-independent, unlike strict membrane dependence binding.
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