Blood coagulation derives from a series of specific proteolytic activation reactions that are catalyzed with narrow and defined specificity by trypsin-like serine proteinases. In several instances, these proteinases function in membrane assembled enzyme complexes. Distinctive protein substrate specificities and the modulation of enzymic function by interactions with membranes and cofactors are hallmarks of the proteolytic reactions of blood coagulation. There are major gaps in the current understanding in the molecular bases for these unique features that underlie the function of the coagulation reactions. This program proposes and integrated approach focused on the contributions of macromolecular interactions to the specificity and modulation of enzymic function that leads to action of the hemostatic enzymes. Project 1 (Krishnaswamy) uses the prothrombinase complex as a paradigm to investigate the role of extended macromolecular interactions between the protein substrate and the enzyme complex in explaining protein substrate specificity and cofactor function. Project 2 (Camire) will investigate the relationship between zymogen proteolysis and the development of discrete macromolecular binding interactions that lead to the assembly of the prothrombinase complex and the expression of enzymic function. Studies are also proposed to assess the role of cofactor-substrate interactions in determining function. Project 3 (Zheng) investigates the link between coagulation and von Willebrand Factor processing by investigating the regulation of ADAMTS13 function by factor VIII and shear stress. The objectives of the three projects will be supported by an administrative core and a core that provides support for molecular biology and protein expression. Overall, this project applies the expertise of the individual investigators towards addressing major unanswered questions in hemostasis. The proposed approaches will provide new insights into the chemistry and biology of the blood coagulation reactions with implications for an understanding of normal hemostasis and thrombosis. Aberrations in the blood clotting system can lead to too much bleeding or excessive clotting. Disorders in the clotting process represent a major cause of disease and death in humans. This program seeks to develop new information by which key reactions of blood clotting are regulated. This information will lead to new concepts and strategies for the treatment of blood clotting-related human disease.

Public Health Relevance

Aberrations in the blood clotting system can lead to too much bleeding or excessive clotting. Disorders in the clotting process represent a major cause of disease and death in humans. This program seeks to develop new information by which key reactions of blood clotting are regulated. This information will lead to new concepts and strategies for the treatment of blood clotting-related human disease.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Program Projects (P01)
Project #
5P01HL074124-10
Application #
8450255
Study Section
Special Emphasis Panel (ZHL1-PPG-J (F2))
Program Officer
Link, Rebecca P
Project Start
2003-08-15
Project End
2014-03-31
Budget Start
2013-04-01
Budget End
2014-03-31
Support Year
10
Fiscal Year
2013
Total Cost
$1,544,150
Indirect Cost
$605,457
Name
Children's Hospital of Philadelphia
Department
Type
DUNS #
073757627
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Thalji, Nabil K; Camire, Rodney M (2017) Rendering factor Xa zymogen-like as a therapeutic strategy to treat bleeding. Curr Opin Hematol 24:453-459
Vadivel, Kanagasabai; Kumar, Yogesh; Bunce, Matthew W et al. (2017) Interaction of factor V B-domain acidic region with its basic region and with TFPI/TFPI2: Structural insights from molecular modeling studies. Int Biol Rev 1:
Thalji, Nabil K; Ivanciu, Lacramioara; Davidson, Robert et al. (2016) A rapid pro-hemostatic approach to overcome direct oral anticoagulants. Nat Med 22:924-32
Bradford, Harlan N; Krishnaswamy, Sriram (2016) The Fragment 1 Region of Prothrombin Facilitates the Favored Binding of Fragment 12 to Zymogen and Enforces Zymogen-like Character in the Proteinase. J Biol Chem 291:11114-23
Ostertag, Eric M; Bdeir, Khalil; Kacir, Stephen et al. (2016) ADAMTS13 autoantibodies cloned from patients with acquired thrombotic thrombocytopenic purpura: 2. Pathogenicity in an animal model. Transfusion 56:1775-85
Liang, Hai Po H; Kerschen, Edward J; Basu, Sreemanti et al. (2015) Coagulation factor V mediates inhibition of tissue factor signaling by activated protein C in mice. Blood 126:2415-23
Pickens, Brandy; Mao, Yingying; Li, Dengju et al. (2015) Platelet-delivered ADAMTS13 inhibits arterial thrombosis and prevents thrombotic thrombocytopenic purpura in murine models. Blood 125:3326-34
Zheng, X Long (2015) ADAMTS13 and von Willebrand factor in thrombotic thrombocytopenic purpura. Annu Rev Med 66:211-25
Baroni, M; Pavani, G; Pinotti, M et al. (2015) Asymmetric processing of mutant factor X Arg386Cys reveals differences between intrinsic and extrinsic pathway activation. Biochim Biophys Acta 1854:1351-6
George, L A; Thalji, N K; Raffini, L J et al. (2015) Correction of human hemophilia A whole blood abnormalities with a novel bypass agent: zymogen-like FXa(I16L). J Thromb Haemost 13:1694-8

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