II GLYCOLIPID BIOSYNTHESIS IN MOUSE T- AND B-LYMPHOMAS

这项研究的直接目标有五个： 1.G1cNAcT-2的分离、鉴定和纯化 (UDP-G1cNAc:nLcOse4Cer-(Beta1-3)-N-acetylglucosaminyltransferase)和 G1cNAcT-3(UDP-G1cNAc:i-core(Beta1-6)N-acetylglucosaminyltransferase) 存在于P-1798的中性、洗涤剂增溶的超级纳盐中 小鼠T淋巴瘤(雌激素诱导)。 2.Ga1T-4的纯化及动力学研究 来自0.1%Triton的(UDP-Gal:LcOse3Cer(Beta1-4)galactosyltransferase) Cf-54-P-1798小鼠T淋巴瘤上清液组分。 3.完成I/I双天线结构的体外生物合成 从先前合成的I/I核心糖脂，使用纯化的GalT-4和 G1cNAcT-2。 4.Fuct-2的特性 (GDP-Fuc:nLcOse4Cer(Alpha1-2)fucosyltransferase)和FUCT-3 贝塔淋巴瘤的(GDP-Fuc:LcOse3Cer(Alpha1-3)fucosyltransferase)(TEPC-15 烷烃诱导和ABLS-140 Abelson病毒诱导)。的决定 生物合成产品中的精确化学键 (FucAlpha-Gal-G1cNAc-Gal-G1c-Cer和FucAlpha-G1cNAc-Gal-G1c-Cer) 来自TEPC-15和ABLS-140。 5.部分Eco RI小鼠基因组文库的筛选 ~(32)P标记的AJ小鼠基因组DNA(Charon 4a中的猎枪)的消化 GALT-4基因的插入片段。翻译产品的间接测定法 使用放射性碘标记的单抗，获得特定的GalT-4 DNA序列 对阵高尔特-4。 我们的总体目标是加深对 1例T淋巴瘤小鼠淋巴网状系统的糖基转移酶 (P-1798)和两个B淋巴瘤(TEPC-15和ABLS-140)作为模型系统。

The immediate objectives of the research are fivefold: 1. Separation, characterization, and purification of G1cNAcT-2 (UDP-G1cNAc:nLcOse4Cer-(Beta1-3)-N-acetylglucosaminyltransferase) and G1cNAcT-3 (UDP-G1cNAc:i-core(Beta1-6)N-acetylglucosaminyltransferase) present in the neutral, detergent-solubilized super natant from p-1798 mouse T-lymphoma (estrogen-induced). 2. Purification and kinetic studies of Ga1T-4 (UDP-Gal:LcOse3Cer(Beta1-4)galactosyltransferase) from the 0.1% Triton CF-54-solubilized supernatant fraction of p-1798 mouse T-lymphoma. 3. Completion of the biosynthesis in vitro of the I/i biantennary structure from previously synthesized I/i core glycolipid, using purified GalT-4 and G1cNAcT-2. 4. Characterization of FucT-2 (GDP-Fuc:nLcOse4Cer(Alpha1-2)fucosyltransferase) and FucT-3 (GDP-Fuc:LcOse3Cer(Alpha1-3)fucosyltransferase) from Beta-lymphoma (TEPC-15 alkane-induced, and ABLS-140 Abelson virus-induced). Determination of exact chemical linkages in the biosynthetic products (FucAlpha-Gal-G1cNAc-Gal-G1c-Cer and FucAlpha-G1cNAc-Gal-G1c-Cer) obtained from TEPC-15 and ABLS-140. 5. Screening of the mouse genomic library, prepared from a partial Eco RI digestion of AJ mouse genomic DNA (Shotgun in Charon 4a) with a 32P-labeled cDNA insert of GalT-4. Indirect assays for the translation products for particular GalT-4 DNA sequences, using radioiodinated monoclonal antibodies against GalT-4. Our overall goal is to develop an understanding of the specificities of glycosyltransferases in mouse lymphoreticular systems using one T-lymphoma (P-1798) and two B-lymphomas (TEPC-15 and ABLS-140) as model systems.