Lactosaminoglycan proteins are a group of glycoproteins present in plasma membranes. Their carbohydrates often serve as cell surface antigens, including Ii, ABH and SSEA-1, which change drastically during cellular differentiation, embryonal development and oncogenesis. Human embryonal carcinoma cell, PA1, resembles the cells of the preblastocyst or trophectoderm of the early blastocyst stage of mouse embryo in many ways. We have determined that large glycopeptides present in PA1 cells are branched lactosaminoglycans with novel disialosyl (NeuNAc alpha2 yields 9 NeuNAc) sequences. We plan to prepare monoclonal antibodies against the NeuNAc alpha2 yields 9 NeuNAc structure to study the distribution of this structure among embryonal cells and tumor cells. Such monoclonal antibodies with defined specificity would also be useful reagents to detect oncodifferentiation antigen and to diagnose tumors. We plan to elucidate the relationship between lactosaminoglycans and complex type oligosaccharides, focusing on their core structures, distribution of lactosaminyl side chains and the occurrence of disialyl structure. PA1 lactosaminoglycans are specifically carried by a glycoprotein (Gp95). Gp95 will be purified and antibodies specific to Gp95 will be prepared. We will examine the distribution of Gp95 among various cell lines by using the antibodies in order to evaluate this glycoprotein as a protein specific to human embryonal carcinoma cells as well as early embryonic cells. We have characterized glycolipids present in PA1 cells and have found that PA1 cells contain mono- and disialylated glycolipids with blood group type 1 sequence. We plan to characterize 0-glycosidic carbohydrates so that the overall structural features of cell surface glycoconjugates on PA1 cells can be established. These studies will provide us with structural information on carbohydrate antigens expressed in teratocarcinoma cells and early stage embryos. Our study will ultimately be helpful for understanding the role of cell surface glycoconjugates in cellular differentiation and oncogenesis.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA034014-06
Application #
3171746
Study Section
Pathobiochemistry Study Section (PBC)
Project Start
1983-02-01
Project End
1990-11-30
Budget Start
1989-01-01
Budget End
1990-11-30
Support Year
6
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Sanford-Burnham Medical Research Institute
Department
Type
DUNS #
009214214
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Fukuda, M N; Masri, K A; Dell, A et al. (1990) Incomplete synthesis of N-glycans in congenital dyserythropoietic anemia type II caused by a defect in the gene encoding alpha-mannosidase II. Proc Natl Acad Sci U S A 87:7443-7
Aoki, D; Appert, H E; Johnson, D et al. (1990) Analysis of the substrate binding sites of human galactosyltransferase by protein engineering. EMBO J 9:3171-8
Fukuda, M N; Masri, K A; Dell, A et al. (1989) Defective glycosylation of erythrocyte membrane glycoconjugates in a variant of congenital dyserythropoietic anemia type II: association of low level of membrane-bound form of galactosyltransferase. Blood 73:1331-9
Masri, K A; Appert, H E; Fukuda, M N (1988) Identification of the full-length coding sequence for human galactosyltransferase (beta-N-acetylglucosaminide: beta 1,4-galactosyltransferase). Biochem Biophys Res Commun 157:657-63
Fukuda, M N; Dell, A; Scartezzini, P (1987) Primary defect of congenital dyserythropoietic anemia type II. Failure in glycosylation of erythrocyte lactosaminoglycan proteins caused by lowered N-acetylglucosaminyltransferase II. J Biol Chem 262:7195-206
Howard, D R; Fukuda, M; Fukuda, M N et al. (1987) The GDP-fucose:N-acetylglucosaminide 3-alpha-L-fucosyltransferases of LEC11 and LEC12 Chinese hamster ovary mutants exhibit novel specificities for glycolipid substrates. J Biol Chem 262:16830-7
Fukuda, M N; Dell, A; Tiller, P R et al. (1986) Structure of a novel sialylated fucosyl lacto-N-norhexaosylceramide isolated from chronic myelogenous leukemia cells. J Biol Chem 261:2376-83
Izhar, M; Siebert, P D; Oshima, R G et al. (1986) Trophoblastic differentiation of human teratocarcinoma cell line HT-H1. Dev Biol 116:510-8
Fukuda, M N; Bothner, B; Lloyd, K O et al. (1986) Structures of glycosphingolipids isolated from human embryonal carcinoma cells. The presence of mono- and disialosyl glycolipids with blood group type 1 sequence. J Biol Chem 261:5145-53
Fukuda, M N; Klier, G; Yu, J et al. (1986) Anomalous clustering of underglycosylated band 3 in erythrocytes and their precursor cells in congenital dyserythropoietic anemia type II. Blood 68:521-9

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