Mannose-containing glycolipids which are sensitive to phosphatidyl-inositol (PI) specific phospholipases are found in eukaryotic cells both as free mannolipids (GPI-lipids) and as anchor moieties of selected membrane proteins (GPI-anchored proteins). Recent research from these and other laboratories has strongly indicated that certain of the free GPI-lipids are precursors of the GPI anchors and that lesions which interrupt their synthesis may account for the lack of expression of GPI-anchored proteins on murine Thy-1 negative T lymphoma mutants and on circulating red and white cells in the hemolytic anemia, Paroxysmal Nocturnal Hemoglobinuria (PNH). The proposed biochemical, cell and molecular biological studies will examine the structures and biosynthesis of both free and protein-associated GPI-lipids, vesicular transport of GPI-lipids and the possible role(s) of GPI-lipids an enzyme substrates and as sources of second messengers. In order to further elucidate the path of GPI-lipid biosynthesis, studies will focus especially on available and new human and murine cell mutants which do not express GPI-anchored proteins, including cells of patients suffering from Zellweger syndrome, which is characterized by deficient synthesis of ether lipids. Related studies will use stable reincorporation of GPI- anchored proteins into the plasma membrane of wild type cells to initiate a line of """"""""cell therapy"""""""", add GPI-lipids onto the surface of mutant cells in an attempt to restore their ability to express GPI-anchored proteins, and clone genes which account for the lesions in GPI-lipid synthesis which are responsible for the lack of cell surface expression of such proteins in PNH.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Program Projects (P01)
Project #
2P01DK038181-06
Application #
3095468
Study Section
Diabetes, Endocrinology and Metabolic Diseases B Subcommittee (DDK)
Project Start
1987-09-01
Project End
1997-08-31
Budget Start
1992-09-30
Budget End
1993-08-31
Support Year
6
Fiscal Year
1992
Total Cost
Indirect Cost
Name
Case Western Reserve University
Department
Type
Schools of Medicine
DUNS #
077758407
City
Cleveland
State
OH
Country
United States
Zip Code
44106
Chen, R; Knez, J J; Merrick, W C et al. (2001) Comparative efficiencies of C-terminal signals of native glycophosphatidylinositol (GPI)-anchored proproteins in conferring GPI-anchoring. J Cell Biochem 84:68-83
Wongkajornsilp, A; Sevlever, D; Rosenberry, T L (2001) Metabolism of exogenous sn-1-alkyl-sn-2-lyso-glucosaminyl-phosphatidylinositol in HeLa D cells: accumulation of glucosaminyl(acyl)phosphatidylinositol in a metabolically inert compartment. Biochem J 359:305-13
Premkumar, D R; Fukuoka, Y; Sevlever, D et al. (2001) Properties of exogenously added GPI-anchored proteins following their incorporation into cells. J Cell Biochem 82:234-45
Sevlever, D; Pickett, S; Mann, K J et al. (1999) Glycosylphosphatidylinositol-anchor intermediates associate with triton-insoluble membranes in subcellular compartments that include the endoplasmic reticulum. Biochem J 343 Pt 3:627-35
Chen, A; Meyerson, H J; Salvekar, A et al. (1998) Non-glycosylated human B7-1(CD80) retains the capacity to bind its counter-receptors. FEBS Lett 428:127-34
Chen, R; Walter, E I; Parker, G et al. (1998) Mammalian glycophosphatidylinositol anchor transfer to proteins and posttransfer deacylation. Proc Natl Acad Sci U S A 95:9512-7
Kraus, D; Medof, M E; Mold, C (1998) Complementary recognition of alternative pathway activators by decay-accelerating factor and factor H. Infect Immun 66:399-405
Sevlever, D; Schiemann, D; Guidubaldi, J et al. (1997) Accumulation of glucosaminyl(acyl)phosphatidylinositol in an S3 HeLa subline expressing normal dolicholphosphomannose synthase activity. Biochem J 321 ( Pt 3):837-44
Yu, J; Nagarajan, S; Knez, J J et al. (1997) The affected gene underlying the class K glycosylphosphatidylinositol (GPI) surface protein defect codes for the GPI transamidase. Proc Natl Acad Sci U S A 94:12580-5
Meyerson, H J; Huang, J H; Fayen, J D et al. (1996) Functional dissociation of CD8 alpha's Ig homologue and connecting peptide domains. J Immunol 156:574-84

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