The major WGA-binding glycoprotein was isolated from the spent culture medium of B16-F10 mouse melanoma cells. A molecular weight of 48,000 was estimated by SDS polyacrylamide gel electrophoresis. Serine and threonine comprised 25% of the total amino acid residues of the glycoprotein. The glycoprotein resembles glycophorin in that it contains both N- and O-linked oligosaccharides, and the O-linked units appear to be in tetrasaccharide structures and clustered on the peptide backbone. Neuraminidase treatment released approximately 80% of the glycoprotein's sialic acid content, and the desialylated species was bound ricin-Sepharose. The interactions of fibronectin with unfractionated histones and histone subfractions (H1, H2A + H3, H2B, and H4) were studied under physiological conditions by laser nephelometry. It was found that fibronectin interacted preferentially with histone H2A + H3. Due to self-aggregation, fibronectin showed a very low relative light scattering. Eleven mouse monoclonal antibodies directed against epitopes on CNBr-peptides of the major sialoglycoconjugate of the human red blood cell, glycophorin A, were produced by hybridomas derived from P3-X63-Ag8.563 myeloma cells and spleen cells from BALB/c mice immunized with purified glycophorin. The monoclonal antibodies could be divided into four groups according to their reactivities with CNBr peptides in a direct ELISA assay: (1) one antibody (6B5) that binds solely to the aminoterminal octapeptide (CNBr3); (2) two antibodies (8F10 and 9C3) that bind to CNBr1 (residues 9-80); (3) two antibodies (3D2 and 4C6) that are reactive with CNBr2, the C-terminal portion of the molecule (residues 81-131); and (4) six antibodies which are cross-reactive with an epitope on both CNBr1 and CNBr3 glycopeptides. This cross-reactive epitope(s) appears to involve both carbohydrate and protein residues. (A)

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA015483-20
Application #
3164190
Study Section
Physiological Chemistry Study Section (PC)
Project Start
1978-06-01
Project End
1988-05-31
Budget Start
1987-06-01
Budget End
1988-05-31
Support Year
20
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Pennsylvania State University
Department
Type
Schools of Medicine
DUNS #
129348186
City
Hershey
State
PA
Country
United States
Zip Code
17033
Aina, Olulanu H; Liu, Ruiwu; Sutcliffe, Julie L et al. (2007) From combinatorial chemistry to cancer-targeting peptides. Mol Pharm 4:631-51
Murray, M C; Bhavanandan, V P; Davidson, E A et al. (1989) Modification of sialyl residues of glycoconjugates by reductive amination. Characterization of the modified sialic acids. Carbohydr Res 186:255-65
Davidson, E A; Perkins, M E (1988) Receptor binding domain of glycophorin A for Plasmodium falciparum surface proteins. Indian J Biochem Biophys 25:90-4
Taddei-Peters, W C; Bhavanandan, V P; Davidson, E A (1988) Purification and partial characterization of a malignancy-associated glycoprotein. Carbohydr Res 182:135-47
Pinnaduwage, P D; Bhavanandan, V P; Davidson, E A (1986) Isolation and characterization of a wheat germ agglutinin-binding glycoprotein from B16 mouse melanoma cells. Carbohydr Res 151:51-64
Pinnaduwage, P D; Bhavanandan, V P; Davidson, E A (1986) Characteristics of two wheat germ agglutinin-resistant variants of B16 mouse melanoma cells with reduced tumorigenicity. Carbohydr Res 151:37-50
Gowda, D C; Bhavanandan, V P; Davidson, E A (1986) Isolation and characterization of proteoglycans secreted by normal and malignant human mammary epithelial cells. J Biol Chem 261:4926-34
Gowda, D C; Bhavanandan, V P; Davidson, E A (1986) Structures of O-linked oligosaccharides present in the proteoglycans secreted by human mammary epithelial cells. J Biol Chem 261:4935-9
Furukawa, K; Minor, J E; Hegarty, J D et al. (1986) Interaction of sialoglycoproteins with wheat germ agglutinin-sepharose of varying ratio of lectin to Sepharose. Use for the purification of mucin glycoproteins from membrane extracts. J Biol Chem 261:7755-61
Poola, I; Seshadri, H S; Bhavanandan, V P (1986) Purification and saccharide-binding characteristics of a rice lectin. Carbohydr Res 146:205-17

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