Stable expression of a cDNA encoding a human beta 1 --> 3galactosyltransferase responsible for lacto-series type 1 core chain synthesis in non-expressing cells: variation in the nature of cell surface antigens expressed.
Stable expression of a cDNA encoding a human beta 1 --> 3galactosyltransferase responsible for lacto-series type 1 core chain synthesis in non-expressing cells: variation in the nature of cell surface antigens expressed.
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编码人β1->3半乳糖基转移酶的cDNA在非表达细胞中稳定表达，该酶负责乳系列1型核心链的合成：表达的细胞表面抗原性质的变化。
DOI：
10.1002/jcb.240500207
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发表时间：
1992
影响因子：
4
通讯作者：
Holmes,EH
中科院分区：
文献类型：
--
作者：
Sherwood,AL;Greene,TG;Holmes,EH
人结肠腺癌科洛205细胞来源的cDNA在通常仅表达新乳糖系列糖脂的细胞系中的瞬时表达导致β1 → 3半乳糖基转移酶基因的表达，该基因负责基于乳糖系列1型核心链合成糖脂。在用1型链末端结构特异性单克隆抗体（TE-3）和与多种1型链衍生物反应的非常广泛特异性抗体（TE-2）的组合进行初始处理后，在抗IgM包被的平板上淘选磷酸钙转染的细胞。淘选后的贴壁细胞能够有效地将β1 → 3-键中的Gal转移至受体糖脂Lc 3。使用这些试剂，产生并分离稳定转染的人结肠腺癌HCT-15细胞的克隆。亲本HCT-15细胞不表达1型链抗原。通过固相抗体结合测定分析在这些克隆的每一个中产生的基于1型链的抗原的性质。观察到三种类型的行为。1型末端结构的形成，要么是专门的唾液酸化或岩藻糖基化，或唾液酸化和岩藻糖基化的决定簇的混合物发生。相反，没有观察到任何克隆和亲本细胞之间的2型抗原表达的差异。这些数据表明，能够修改1型链结构的后续反应的协调在所有克隆中是不相同的。这些结果的碳水化合物生物合成的细胞调节方面的关系进行了讨论。© 1992 Wiley利斯公司
Transient expression of a human colonic adenocarcinoma Colo 205 cell derived cDNA in cell lines which ordinarily express only neolacto‐series glycolipids has resulted in the expression of a β1 → 3galactosyltransferase gene responsible for synthesis of glycolipids based upon the lacto‐series type 1 core chain. Calcium phosphate transfected cells were panned on anti‐lgM coated plates after initial treatment with a combination of monoclonal antibodies specific for type 1 chain terminal structures (TE‐3) and a very broadly specific antibody reactive with multiple type 1 chain derivatives (TE‐2). Adherent cells after panning were capable of efficiently transferring Gal in β1 → 3‐linkage to the acceptor glycolipid Lc3. Using these reagents, clones of stably transfected human colonic adenocarcinoma HCT‐15 cells were produced and isolated. Parental HCT‐15 cells do not express type 1 chain based antigens. The nature of the type 1 chain based antigens produced in each of these clones was analyzed by solid phase antibody binding assays. Three types of behavior were observed. Formation of type 1 terminal structures that were either exclusively sialylated or fucosylated, or a mixture of sialylated and fucosylated determinants occurred. In contrast, no difference in type 2 antigen expression between any clone and the parental cells was observed. These data suggest that coordination of subsequent reactions capable of modifying type 1 chain structures is not the same in all clones. The relationship of these results to aspects of cellular regulation of carbohydrate biosynthesis is discussed. © 1992 Wiley‐Liss, Inc.