The LONG-TERM goal of our research is to study and analyze the structure and function of the factor V molecule in order to understand key mechanisms involved in maintaining hemostasis in the human body. In order to understand these functions, it is necessary to implement the SHORT-TERM goal of identifying the amino acid residue(s) of factor V that interact with the blood coagulation factor prothrombin (flla), during prothrombinase complex assembly and function.
The specific aims of the project are stated as follows: 1) To identify the specific amino acid residue(s) of the carboxyl-terminus of the factor Va heavy chain that interact with prothrombin during prothrombinase complex formation. 2) To identify the individual amino acid(s) of factor Va that interact with prothrombin and factor Xa required to drive meizothrombin formation. 3) To identify specific acidic amino acid(s) of the factor Va heavy chain that are involved in prothrombinase complex formation. To obtain our data, factor V mutants will be constructed by PCR-based methods and purified plasmids will be transiently transfected into COS-7L cells in order to obtain recombinant proteins that will be anayzed by SDS-PAGE analysis and Western Blotting. Proteins will be screened for clotting activity in one-and two-stage clotting assays. Kinetic analyses will be done by measuring thrombin formation. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Predoctoral Individual National Research Service Award (F31)
Project #
5F31HL085928-02
Application #
7354838
Study Section
Special Emphasis Panel (ZRG1-DIG-H (29))
Program Officer
Sarkar, Rita
Project Start
2007-02-01
Project End
2011-01-31
Budget Start
2008-02-01
Budget End
2009-01-31
Support Year
2
Fiscal Year
2008
Total Cost
$30,967
Indirect Cost
Name
Cleveland State University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
010841617
City
Cleveland
State
OH
Country
United States
Zip Code
44115
Hirbawi, Jamila; Vaughn, John L; Bukys, Michael A et al. (2010) Contribution of amino acid region 659-663 of Factor Va heavy chain to the activity of factor Xa within prothrombinase . Biochemistry 49:8520-34
Hirbawi, Jamila; Bukys, Michael A; Barhoover, Melissa A et al. (2008) Role of the acidic hirudin-like COOH-terminal amino acid region of factor Va heavy chain in the enhanced function of prothrombinase. Biochemistry 47:7963-74