Abstract

Possible relationships between tumorigenicity and expression of beta-GalNAcT-1, beta-GalNAcT-2, and alpha-GalNAcT-3 involved in Gg- and Gb-core-containing glycosphingolipid biosynthesis and SAT-3 in LM1 biosynthesis will be a major emphasis of this research proposal. The overall immediate objectives of this research program are to solubilize and purify these three N-acetylgalactosaminyltransferases from guinea pig tumor cells (104Cl and 106B) and embryonic chicken brains by anion exchange and specific affinity chromatography. HPLC of ?125?I-GSLs (from cell surfaces) and enzymatic studies will also be carried out to compare the synthesis of Forssman antigen (glycolipid) during the growth cycle of guinea pig 104Cl and 106B cultured cells. Radioactive products will be isolated from [?14?C-Ac]-GbOse4Cer and [?3?H]GbOse3Cer after reaction with unlabeled UDP-GalNAc. The exact chemical linkages of the terminal GalNAc residues will be determined after permethylation of purified enzymatic products and characterization of the partially methylated radioactive [?3?H]galactose and [?14?C-Ac]GalNAc units obtained by autoradiography. Anomeric linkages will be determined by using highly purified beta-hexosaminidase (jack bean and papaya) and alpha-hexosaminidase (clam). Purification of alpha-hexosaminidase from clam is also part of this project. Differential inhibition of these three GalNAct activities will be studied in the presence of UDP, IDP, pCMB, and NEM. Biosynthesis in vitro of LM1 (sialosyl-neolactotetraosylceramide; NeuAc-nLcOse4Cer) in mouse N-18 clone and in human neuroblastoma 1MR-32 cells will be investigated in the third year of proposed research. Binding of [?125?I]-labeled GSLs, lectin (Eunonymus europeus) and toxins (cholera and ricin) to neuroblastoma cell surfaces will also be studied before and after chemically induced differentiation with (But)?2? cAMP and HMBA. (A)

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA014764-09
Application #
3164007
Study Section
Pathobiochemistry Study Section (PBC)
Project Start
1979-04-01
Project End
1986-09-30
Budget Start
1985-04-01
Budget End
1986-09-30
Support Year
9
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
University of Notre Dame
Department
Type
Schools of Arts and Sciences
DUNS #
824910376
City
Notre Dame
State
IN
Country
United States
Zip Code
46556

  Publications

Basu, Subhash; Ma, Rui; Moskal, Joseph R et al. (2012) Ganglioside biosynthesis in developing brains and apoptotic cancer cells: X. regulation of glyco-genes involved in GD3 and Sialyl-Lex/a syntheses. Neurochem Res 37:1245-55
Ma, Rui; Hopp, Elizabeth A; Decker, N Matthew et al. (2011) Regulation of glycosyltransferase genes in apoptotic breast cancer cells induced by L: -PPMP and cisplatin. Adv Exp Med Biol 705:621-42
Ma, Rui; Matthew Decker, N; Anilus, Vesta et al. (2009) Post-translational and transcriptional regulation of glycolipid glycosyltransferase genes in apoptotic breast carcinoma cells: VII. Studied by DNA-microarray after treatment with L-PPMP. Glycoconj J 26:647-61
Ray, S; Kelley, T J; Fan, L et al. (1994) Characterization of DNA polymerase-alpha/primase complex from developing embryonic chicken brains. Indian J Biochem Biophys 31:226-35
Kelley, T J; Moghaddas, S; Bose, R et al. (1993) Inhibition of immunopurified DNA polymerase-alpha from PA-3 prostate tumor cells by platinum (II) antitumor drugs. Cancer Biochem Biophys 13:135-46
Basu, M; Basu, S S; Li, Z et al. (1993) Biosynthesis and regulation of Le(x) and SA-Le(x) glycolipids in metastatic human colon carcinoma cells. Indian J Biochem Biophys 30:324-32
Schaeper, R J; Das, K K; Li, Z et al. (1992) In vitro biosynthesis of GbOse4Cer (globoside) and GM2 ganglioside by the (1-->3) and (1-->4)-N-acetyl beta-D-galactosaminyltransferases from embryonic chicken brain. Solubilization, purification, and characterization of the transferases. Carbohydr Res 236:227-44
Basu, M; Hawes, J W; Li, Z et al. (1991) Biosynthesis in vitro of SA-Lex and SA-diLex by alpha 1-3 fucosyltransferases from colon carcinoma cells and embryonic brain tissues. Glycobiology 1:527-35
Basu, M; Khan, F A; Das, K K et al. (1991) Biosynthesis in vitro of core lacto-series glycosphingolipids by N-acetyl-D-glucosaminyltransferases from human colon carcinoma cells, Colo 205. Carbohydr Res 209:261-77
Ray, S; Kelley, T J; Campion, S et al. (1991) Developmental expression of the embryonic chicken brain DNA polymerase alpha and its binding with monoclonal antibodies against human KB cell DNA polymerase alpha. Cell Growth Differ 2:567-73

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