Abstract

There are three different though related objectives of the proposed studies. First, we plan to investigate the binding characteristics of platelet-activating factor to both intact cells and membrane preparation, the role of the divalent cation, calcium, and the biochemical changes that are coupled to each receptor population. Second, we wish to elucidate the mechanism(s) by which phorbol 12-myristae 13-acetate inhibits the action of the platelet-activating factor and the role of protein kinase C in this inhibition. These include measuring the effects of the phorbol esters on: binding of platelet-activating factor, phosphoinositide hydrolysis, the dissociation of the G-protein, GTPase activities, and binding of nonhydrolysable GTP analogues. Third, we plan to characterize the effects of calcium channel blockers on binding of platelet-activating factor and the biochemical changes it produces.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL053786-08
Application #
2231893
Study Section
Lung Biology and Pathology Study Section (LBPA)
Project Start
1987-09-30
Project End
1998-07-31
Budget Start
1996-08-01
Budget End
1998-07-31
Support Year
8
Fiscal Year
1996
Total Cost
Indirect Cost

  Institution

Name
University of Connecticut
Department
Anatomy/Cell Biology
Type
Schools of Dentistry
DUNS #
City
Farmington
State
CT
Country
United States
Zip Code
06030

  Publications

Syrbu, S I; Waterman, W H; Molski, T F et al. (1999) Phosphorylation of cytosolic phospholipase A2 and the release of arachidonic acid in human neutrophils. J Immunol 162:2334-40
Zu, Y L; Qi, J; Gilchrist, A et al. (1998) p38 mitogen-activated protein kinase activation is required for human neutrophil function triggered by TNF-alpha or FMLP stimulation. J Immunol 160:1982-9
Zu, Y L; Ai, Y; Gilchrist, A et al. (1997) High expression and activation of MAP kinase-activated protein kinase 2 in cardiac muscle cells. J Mol Cell Cardiol 29:2159-68
Waterman, W H; Molski, T F; Huang, C K et al. (1996) Tumour necrosis factor-alpha-induced phosphorylation and activation of cytosolic phospholipase A2 are abrogated by an inhibitor of the p38 mitogen-activated protein kinase cascade in human neutrophils. Biochem J 319 ( Pt 1):17-20
Nahas, N; Waterman, W H; Sha'afi, R I (1996) Granulocyte-macrophage colony-stimulating factor (GM-CSF) promotes phosphorylation and an increase in the activity of cytosolic phospholipase A2 in human neutrophils. Biochem J 313 ( Pt 2):503-8
Zu, Y L; Ai, Y; Gilchrist, A et al. (1996) Activation of MAP kinase-activated protein kinase 2 in human neutrophils after phorbol ester or fMLP peptide stimulation. Blood 87:5287-96
Waterman, W H; Sha'afi, R I (1995) A mitogen-activated protein kinase independent pathway involved in the phosphorylation and activation of cytosolic phospholipase A2 in human neutrophils stimulated with tumor necrosis factor-alpha. Biochem Biophys Res Commun 209:271-8
Waterman, W H; Sha'afi, R I (1995) Effects of granulocyte-macrophage colony-stimulating factor and tumour necrosis factor-alpha on tyrosine phosphorylation and activation of mitogen-activated protein kinases in human neutrophils. Biochem J 307 ( Pt 1):39-45
Fouda, S I; Molski, T F; Ashour, M S et al. (1995) Effect of lipopolysaccharide on mitogen-activated protein kinases and cytosolic phospholipase A2. Biochem J 308 ( Pt 3):815-22