ptFVa ( Pseudonaja Textilis Venom-Derived Factor Va) Retains Structural Integrity Following Proteolysis by Activated Protein C

• Published in: Arteriosclerosis, thrombosis, and vascular biology Vol. 41; no. 8; pp. 2263 - 2276
• Main Authors: Schreuder, Mark, Liu, Xiaosong, Cheung, Ka Lei, Reitsma, Pieter H., Nicolaes, Gerry A.F., Bos, Mettine H.A.
• Format: Journal Article
• Language: English
• Published: United States Lippincott Williams & Wilkins 01.08.2021
• Subjects: Animals; Basic Sciences; Cell Line; Cricetinae; Elapid Venoms - chemistry; Elapid Venoms - metabolism; Enzyme Activation; Factor Va - chemistry; Factor Va - genetics; Factor Va - metabolism; Humans; Hydrogen Bonding; Molecular Dynamics Simulation; Protein C - metabolism; Protein Interaction Domains and Motifs; Proteolysis; Structure-Activity Relationship; Substrate Specificity; activated protein C resistance; snake venoms; blood coagulation; factor V
• ISSN: 1079-5642; 1524-4636
• DOI: 10.1161/ATVBAHA.121.316038

OBJECTIVE: The Australian snake venom ptFV (Pseudonaja textilis venom-derived factor V) variant retains cofactor function despite APC (activated protein C)-dependent proteolysis. Here, we aimed to unravel the mechanistic principles by determining the role of the absent Arg306 cleavage site that is required for the inactivation of FVa (mammalian factor Va). APPROACH AND RESULTS: Our findings show that in contrast to human FVa, APC-catalyzed proteolysis of ptFVa at Arg306 and Lys507 does not abrogate ptFVa cofactor function. Remarkably, the structural integrity of APC-proteolyzed ptFVa is maintained indicating that stable noncovalent interactions prevent A2-domain dissociation. Using Molecular Dynamics simulations, we uncovered key regions located in the A1 and A2 domain that may be at the basis of this remarkable characteristic. CONCLUSIONS: Taken together, we report a completely novel role for uniquely adapted regions in ptFVa that prevent A2 domain dissociation. As such, these results challenge our current understanding by which strict regulatory mechanisms control FVa activity.