权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

GLYCOLIPID METABOLISM IN TUMOR AND TRANSFORMED CELLS

肿瘤和转化细胞中的糖脂代谢

基本信息

批准号：
3164011
负责人：
SUBHASH C BASU
金额：
$ 13.39万
依托单位：
UNIVERSITY OF NOTRE DAME
依托单位国家：
美国
项目类别：
财政年份：
1979
资助国家：
美国
起止时间：
1979-04-01 至 1992-06-30
项目状态：
已结题

项目摘要

The immediate objectives of the proposed research are sixfold: 1. Separation, characterization, and purification of GalNAcT-2(UDP-GalNAc:GbOse3Cer(Betal-3)N-acetylgalactosaminyltransferase) and GalNAcT-3(UDP-GalNAc:GbOse4Cer(Alphal-3)N-acetyl-galactosaminyltransferase) present in the detergent solubilized supernatants from guinea pig tumor cell lines 104Cl (tumorigenic) and 106B(nontumorigenic), respectively. 2. Determination of the exact chemical linkages in the Forssman biosynthetic products of the 104Cl and 106B GalNAcT-3-catalyzed reactions. 3. Purification and kinetic studies of GalNAcT-1(UDP-GalNAc:GM3 or Lac-Cer(Betal-4)N-acetyl-galactosaminyltransferase) from guinea pig bone marrow. 4. Determination of the effects of tunicamycin and insulin on GalNAcT-2 and GalNAcT-3 activities in 104Cl and 106B guinea pig tumor cells. 5. Biosynthesis in vitro of the tumor-specific antigens sialosylAlpha2-3Le-x, sialosylAlpha2-3Le-a in neuroblastoma IMR-32 cells and GD3 and Fuc-GM1 in rat prostate PA-I/II cells. 6. Characterization of GalT-5(UDP-Gal:nLcOse4Cer(Alphal-3)galactosyltransferase) from neuroblastoma IMR-32 cells and study of its activity under various growth conditions during the cell cycle and in the presence of differentiating agent ((But)2cAMP and HMBA). Our overall goal is to develop an understanding of the specificities of glycolipid:glycosyltransferases in three tumor cell lines of guinea pig and human origin. Tunicamycin inhibits glycoconjugate biosynthesis. However, inhibition of glycosylation of glycosyltransferase is a novel observation. At the end of this project an attempt will be made to establish a correlation between inhibition of a glycosyltransferase (GalNAcT-2) by tunicamycin and the change of a tumor-specific antigen (Forssman GSL) on the surface of 104Cl and 106B cells using 125I-labeled Helix pomatia and Dolichos biflorus lectins and Siga toxin. Screening of the mouse genomic library, prepared from a partial EcoRI digestion of AJ mouse genomic DNA (Shotgun in Charon 4a) with a 32P-labeled cDNA insert of GalNAcT-2 will be performed. Using 125I-mono and polyclonal antibodies the translated products for particular 104Cl GalNAcT-2 DNA sequences will also be screened.

拟议研究的直接目标有六个： 1. 分离、表征和纯化 GalNAcT-2(UDP-GalNAc:GbOse3Cer(Betal-3)N-乙酰半乳糖氨基转移酶) 和 GalNAcT-3(UDP-GalNAc:GbOse4Cer(Alphal-3)N-乙酰基半乳糖氨基转移酶) 存在于豚鼠肿瘤的去污剂溶解的上清液中分别为细胞系104C1（致瘤性）和106B（非致瘤性）。 2. 福斯曼中确切化学键的确定 104Cl 和 106B GalNAcT-3 催化反应的生物合成产物。 3. GalNAcT-1(UDP-GalNAc:GM3 或 Lac-Cer(Betal-4)N-乙酰基-半乳糖氨基转移酶) 来自豚鼠骨骨髓。 4. 确定衣霉素和胰岛素对 GalNAcT-2 和 GalNAcT-3 在 104Cl 和 106B 豚鼠肿瘤细胞中的活性。 5. 肿瘤特异性抗原的体外生物合成神经母细胞瘤 IMR-32 细胞中的 sialosylAlpha2-3Le-x、sialosylAlpha2-3Le-a 以及大鼠前列腺PA-I/II细胞中的GD3和Fuc-GM1。 6. 表征 GalT-5(UDP-Gal:nLcOse4Cer(Alphal-3)半乳糖基转移酶)来自神经母细胞瘤IMR-32细胞及其在不同生长条件下的活性研究细胞周期期间和存在分化的条件剂（(But)2cAMP 和 HMBA）。我们的总体目标是了解糖脂：豚鼠三种肿瘤细胞系中的糖基转移酶人类起源。 Tunicamycin 抑制复合糖的生物合成。然而，抑制糖基转移酶的糖基化是一个新的观察结果。在该项目结束时，将尝试建立一个糖基转移酶 (GalNAcT-2) 抑制之间的相关性衣霉素和肿瘤特异性抗原（福斯曼 GSL）的变化使用 125I 标记的 Helix pomatia 检测 104Cl 和 106B 细胞的表面双花扁豆凝集素和西加毒素。小鼠基因组筛选文库，由 AJ 小鼠基因组 DNA 的部分 EcoRI 消化制备（Charon 4a 中的霰弹枪）带有 GalNAcT-2 的 32P 标记 cDNA 插入片段将被执行。使用 125I-单克隆和多克隆抗体翻译还将筛选针对特定 104Cl GalNAcT-2 DNA 序列的产品。