Regulation of factor V and factor V-short by TFPI&#945;: Relationship between B-domain proteolysis and binding.

Petrillo, Teodolinda; Ayombil, Francis; Van't Veer, Cornelis; Camire, Rodney M

Regulation of factor V and factor V-short by TFPIα: Relationship between B-domain proteolysis and binding.

Petrillo, Teodolinda; Ayombil, Francis; Van't Veer, Cornelis; Camire, Rodney M.

Afiliación

Petrillo T; Division of Hematology and the Raymond G. Perelman Center for Cellular and Molecular Therapeutics, The Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA.
Ayombil F; Division of Hematology and the Raymond G. Perelman Center for Cellular and Molecular Therapeutics, The Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA.
Van't Veer C; Center of Experimental and Molecular Medicine, Amsterdam UMC, University of Amsterdam, Amsterdam, the Netherlands.
Camire RM; Division of Hematology and the Raymond G. Perelman Center for Cellular and Molecular Therapeutics, The Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA; Department of Pediatrics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA. Electronic

J Biol Chem ; 296: 100234, 2021.

Article en En | MEDLINE | ID: mdl-33376137

RESUMEN

Coagulation factor V (FV) plays an anticoagulant role but serves as a procoagulant cofactor in the prothrombinase complex once activated to FVa. At the heart of these opposing effects is the proteolytic removal of its central B-domain, including conserved functional landmarks (basic region, BR; 963-1008 and acidic region 2, AR2; 1493-1537) that enforce the inactive FV procofactor state. Tissue factor pathway inhibitor α (TFPIα) has been associated with FV as well as FV-short, a physiologically relevant isoform with a shortened B-domain missing the BR. However, it is unclear which forms of FV are physiologic ligands for TFPIα. Here, we characterize the binding and regulation of FV and FV-short by TFPIα via its positively charged C-terminus (TFPIα-BR) and examine how bond cleavage in the B-domain influences these interactions. We show that FV-short is constitutively active and functions in prothrombinase like FVa. Unlike FVa, FV-short binds with high affinity (Kd â¼1 nM) to TFPIα-BR, which blocks procoagulant function unless FV-short is cleaved at Arg1545, removing AR2. Importantly, we do not observe FV binding (µM detection limit) to TFPIα. However, cleavage at Arg709 and Arg1018 displaces the FV BR, exposing AR2 and allowing TFPIα to bind via its BR. We conclude that for full-length FV, the detachment of FV BR from AR2 is necessary and sufficient for TFPIα binding and regulation. Our findings pinpoint key forms of FV, including FV-short, that act as physiologic ligands for TFPIα and establish a mechanistic framework for assessing the functional connection between these proteins.

Asunto(s)

Factor V/química; Factor Va/química; Lipoproteínas/química; Trombina/genética; Coagulación Sanguínea/genética; Factor V/genética; Factor Va/genética; Factor Xa/química; Factor Xa/genética; Humanos; Ligandos; Lipoproteínas/genética; Unión Proteica/genética; Dominios Proteicos/genética; Proteolisis/efectos de los fármacos; Trombina/química; Tromboplastina/química; Tromboplastina/genética

Palabras clave

coagulation factor; factor V; factor Va; hemostasis; protein complex; prothrombin; prothrombinase; thrombin; tissue factor pathway inhibitor

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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Factor V / Trombina / Factor Va / Lipoproteínas Límite: Humans Idioma: En Revista: J Biol Chem Año: 2021 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos

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