Abstract

The objective of this grant is to elucidate basic mechanisms of hemostasis and to devise strategies to alter the natural history of thrombotic disorders. There are two projects. The first deals with the role of platelets, leukocytes, and endothelial cells in the initiation and control of coagulation reactions. Prothrombin activation on platelets will be studied using homogeneous factor Va and its component peptides. The role of factor Va in protein C activation will be elucidated using the isolated factor Va peptides. The relation between thrombomodulin and factor Va, two proteins that both participate in protein C activation, will be studied using cultured endothelial cells and antibodies to thrombomodulin and factor Va. Monoclonal antibodies to protein C, factor Va and its component chains will be prepared. The platelet protease that activates factor V will be purified and its role in the initiation of platelet surface prothrombin activation will be investigated using antibodies specific for factor Va and the western blotting technique. The second project deals with the mechanism of eicosanoid precursor uptake and release by platelets and cultured cells. The fatty acid specificity of arachidonoyl CoA synthetase will be elucidated using a variety of 14C-labeled polyunsaturated fatty acids and compared to the high affinity fatty acid uptake capacity of platelets. The fatty acid specificity of the agonist-induced fatty acid release mechanism of platelets will also be elucidated. Arachidonoyl CoA synthetase will be purified to homogeneity and antibodies prepared. The technique of suicide selection after mutagenesis will be used to select arachidonate uptake and release mutants of HSD mouse fibrosarcoma cells. Mutants will be isolated by cloning and replica-plating on polyester cloth. Mutant cell lines will be useful in the elucidation of mechanisms of fatty acid uptake and release in these cells.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37HL016634-27
Application #
3485504
Study Section
Hematology Subcommittee 2 (HEM)
Project Start
1979-04-01
Project End
1994-03-31
Budget Start
1992-04-01
Budget End
1993-03-31
Support Year
27
Fiscal Year
1992
Total Cost
Indirect Cost

  Institution

Name
Washington University
Department
Type
Schools of Medicine
DUNS #
062761671
City
Saint Louis
State
MO
Country
United States
Zip Code
63130

  Publications

Zhang, Chunfen; Majerus, Philip W; Wilson, Monita P (2012) Regulation of inositol 1,3,4-trisphosphate 5/6-kinase (ITPK1) by reversible lysine acetylation. Proc Natl Acad Sci U S A 109:2290-5
Zou, Jun; Zhang, Chunfen; Marjanovic, Jasna et al. (2012) Myotubularin-related protein (MTMR) 9 determines the enzymatic activity, substrate specificity, and role in autophagy of MTMR8. Proc Natl Acad Sci U S A 109:9539-44
Zou, Jun; Majerus, Philip W; Wilson, David B et al. (2012) The role of myotubularin-related phosphatases in the control of autophagy and programmed cell death. Adv Biol Regul 52:282-9
Ferron, Mathieu; Boudiffa, Maya; Arsenault, Michel et al. (2011) Inositol polyphosphate 4-phosphatase B as a regulator of bone mass in mice and humans. Cell Metab 14:466-77
Marjanovic, Jasna; Wilson, Monita P; Zhang, Chunfen et al. (2011) The role of inositol polyphosphate 4-phosphatase 1 in platelet function using a weeble mouse model. Adv Enzyme Regul 51:101-5
Traynor-Kaplan, Alexis E; Moody, Mark; Nur, Magda et al. (2010) INO-4995 therapeutic efficacy is enhanced with repeat dosing in cystic fibrosis knockout mice and human epithelia. Am J Respir Cell Mol Biol 42:105-12
Majerus, Philip W; Wilson, David B; Zhang, Chunfen et al. (2010) Expression of inositol 1,3,4-trisphosphate 5/6-kinase (ITPK1) and its role in neural tube defects. Adv Enzyme Regul 50:365-72
Jacoby, Monique; Cox, James J; Gayral, Stéphanie et al. (2009) INPP5E mutations cause primary cilium signaling defects, ciliary instability and ciliopathies in human and mouse. Nat Genet 41:1027-31
Bielas, Stephanie L; Silhavy, Jennifer L; Brancati, Francesco et al. (2009) Mutations in INPP5E, encoding inositol polyphosphate-5-phosphatase E, link phosphatidyl inositol signaling to the ciliopathies. Nat Genet 41:1032-6
Zou, Jun; Chang, Shao-Chun; Marjanovic, Jasna et al. (2009) MTMR9 increases MTMR6 enzyme activity, stability, and role in apoptosis. J Biol Chem 284:2064-71

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