Abstract

The long-term goals of this research program are to increase our basic knowledge of how plasma proteins interact with circulating blood platelets leading to their activation and shape change at the site of vascular injury. These reactions play a significant role in cardiovascular disease leading to heart attack and stroke in humans. The approach includes an investigation of the signaling process initiated on the surface of the platelet by thrombin and von Willebrand factor (VWF) and their relationship to the proteins of the cytoskeleton that control the shape of the platelets. These studies will focus on the glycoprotein Ib-lX-V complex, its binding to the a, b, and g filamins in the platelets, and the relationship of the filamins to a guanine nucleotide exchange factor (GEF) with DH-PH domains. This GEF called filamin associated protein has been characterized in our laboratory. These studies will then be extended to the specific small GTPases such as Rho, Rac, and Cdc42 that function as substrates for the GEFs and play a specific role in the development of filopodia, lamellipodia, stress fibers and focal adhesions when platelets are activated. In parallel experiments, the protease-activated receptors (PAR1 and PAR4) will be further examined for their role in platelet shape change via the small GTPases (Rho, Rac, Cdc42) and the platelet GEFs.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
2R01HL016919-30
Application #
6678715
Study Section
Hematology Subcommittee 2 (HEM)
Program Officer
Link, Rebecca P
Project Start
1997-09-01
Project End
2007-08-31
Budget Start
2003-09-01
Budget End
2004-08-31
Support Year
30
Fiscal Year
2003
Total Cost
$424,464
Indirect Cost

  Institution

Name
University of Washington
Department
Biochemistry
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195

  Publications

Black, Peter C; Mize, Gregory J; Karlin, Peter et al. (2007) Overexpression of protease-activated receptors-1,-2, and-4 (PAR-1, -2, and -4) in prostate cancer. Prostate 67:743-56
Kulman, John D; Satake, Masanobu; Harris, Jeff E (2007) A versatile system for site-specific enzymatic biotinylation and regulated expression of proteins in cultured mammalian cells. Protein Expr Purif 52:320-8
Kulman, John D; Harris, Jeff E; Xie, Ling et al. (2007) Proline-rich Gla protein 2 is a cell-surface vitamin K-dependent protein that binds to the transcriptional coactivator Yes-associated protein. Proc Natl Acad Sci U S A 104:8767-72
Davie, Earl W; Kulman, John D (2006) An overview of the structure and function of thrombin. Semin Thromb Hemost 32 Suppl 1:3-15
Kulman, John D; Harris, Jeff E; Nakazawa, Noriko et al. (2006) Vitamin K-dependent proteins in Ciona intestinalis, a basal chordate lacking a blood coagulation cascade. Proc Natl Acad Sci U S A 103:15794-9
Pratt, Kathleen P; Qian, Jiahua; Ellaban, Ekram et al. (2004) Immunodominant T-cell epitopes in the factor VIII C2 domain are located within an inhibitory antibody binding site. Thromb Haemost 92:522-8
Takeshima, Kazuya; Smith, Christina; Tait, Jonathan et al. (2003) The preparation and phospholipid binding property of the C2 domain of human factor VIII. Thromb Haemost 89:788-94
Greenberg, Daniel L; Mize, Gregory J; Takayama, Thomas K (2003) Protease-activated receptor mediated RhoA signaling and cytoskeletal reorganization in LNCaP cells. Biochemistry 42:702-9
Fuentes-Prior, Pablo; Fujikawa, Kazuo; Pratt, Kathleen P (2002) New insights into binding interfaces of coagulation factors V and VIII and their homologues lessons from high resolution crystal structures. Curr Protein Pept Sci 3:313-39
Chung, Dominic W; Fujikawa, Kazuo (2002) Processing of von Willebrand factor by ADAMTS-13. Biochemistry 41:11065-70

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