Glycosphingolipids (GSLs) comprise a diverse class of biomolecules that are known to function as genetically-controlled recognition markers and modulators of membrane receptors and to be the targets of pathogenic agents for many cellular systems. However, very little is known about the GSls of human blood platelets. Thus, we propose, as a long-term objective, to study the pathobiochemistry of platelet GSLs. The molecular structure of all 24 platelet GSLs that we have detected by high-performance thin-layer chromatography (HPTLC) will be studied through: (1) large-scale extraction, fractionation (DEAE-Sephadex, Bio-Sil A), and purification (HPLC), (2) one-and two-dimensional proton nuclear magnetic resonance spectroscopy, (3) fast atom bombardment mass spectrometry, (4) gas chromatography. (5) microscale methylation analysis, (6) HPTLC-immunostaining, and (7) specific glycosidase treatment. GSLs with unusual structure will be used to prepare mono- clonal antibodies. The immunogenetic expression of platelet GSls will be studied by analyzing the HPTlC patterns of GSls found in individuals within a population of normal, HLA-typed donors. The function of platelet GSls will be studied by measuring their effect on ligand binding in three quantitative receptor systems. These binding systems platelet aggregation, (2) platelet serotonin uptake and binding and (3) 125I-labeled fibrinogen, fibronectin or von Willebrand factor binding to purified glycoprotein IIb/IIIa phospholipid vesicles. The role of platelet GSLs as targets for pathogenic agents will be assessed by studying the binding of patient alloantibodies, verotoxin, and E. coli to immobilized GSls using HPTLC immunostaining and HPTlC-autoradiography. The information generated through this project will contribute to a better understanding of the physiology and pathophysiology of: (1) hemostasis and thrombosis, (2) coronary artery vasospasm and hypertension, and (3) thrombocytopenias observed in patients with leukemia, infection, or undergoing bone marrow transplantation.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
5R29HL042395-03
Application #
3472546
Study Section
Pathobiochemistry Study Section (PBC)
Project Start
1989-04-01
Project End
1994-03-31
Budget Start
1991-04-01
Budget End
1992-03-31
Support Year
3
Fiscal Year
1991
Total Cost
Indirect Cost
Name
University of Iowa
Department
Type
Schools of Medicine
DUNS #
041294109
City
Iowa City
State
IA
Country
United States
Zip Code
52242
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Cooling, L L; Zhang, D; Koerner, T A (2001) Human platelets express gangliosides with LKE activity and ABH blood group activity. Transfusion 41:504-16
Cunningham, M T; Citron, B A; Koerner, T A (1999) Evidence of a phospholipid binding species within human fibrinogen preparations. Thromb Res 95:325-34
Cooling, L L; Walker, K E; Gille, T et al. (1998) Shiga toxin binds human platelets via globotriaosylceramide (Pk antigen) and a novel platelet glycosphingolipid. Infect Immun 66:4355-66
Cooling, L L; Zhang, D S; Walker, K E et al. (1995) Detection in human blood platelets of sialyl Lewis X gangliosides, potential ligands for CD62 and other selectins. Glycobiology 5:571-81
Cooling, L L; Koerner, T A; Naides, S J (1995) Multiple glycosphingolipids determine the tissue tropism of parvovirus B19. J Infect Dis 172:1198-205
Cunningham, M T; Olson, J D; Koerner, T A (1993) Glycosphingolipid inhibition of the adhesion of thrombin-activated platelets to surfaces is potentiated by albumin. Glycobiology 3:331-7
Greer, J M; Koerner, T A; Hayakawa, K et al. (1993) The 3G11+ antigen, a marker for murine CD4+ TH1 lymphocytes, is a ganglioside. Glycobiology 3:391-401
Koerner, T A; Cunningham, M T; Zhang, D S (1992) The role of membrane lipid in the platelet storage lesion. Blood Cells 18:481-97;discussion 498-500