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Biosynthesis in vitro of fucose-containing glycosphingolipids in human neuroblastoma IMR-32 cells.

机译:人神经母细胞瘤IMR-32细胞中含岩藻糖的糖鞘脂的体外生物合成。

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摘要

Two different glycolipid:fucosyltransferase activities involved in the biosynthesis in vitro of blood group-related glycosphingolipids have been detected in a membrane preparation isolated from a human neuroblastoma-derived clonal cell line, IMR-32. The membrane preparation contains an alpha (1,2)-fucosyltransferase (EC 2.4.1.89) that catalyzed the transfer of vucose from GDP--[14C]fucose to neolactotetraosylceramide or neolactopentaosylceramide to form types H-I and B-I glycolipids, respectively. The second fucosyltransferase catalyzes the transfer of fucose to lactotriaosylceramide [GlcNAc(beta1-3)Gal(beta1-4)Glc-Cer] to form a tetraglycosylceramide intermediate of the novel Lea-type glycolipid. UDP-galactose:lactotriaosylceramide beta-galactosyltransferase (EC 2.4.1.86) had 4 times the activity of UDP-galactose:alpha-galactosyltransferase (EC 2.4.1.87) when tested under similar conditions. alpha-Fucosyltransferase activities and the incorporation of [14C]fucose into glycoproteins and glycolipids were also compared in cells differentiated in the presence of 4 micron BrdUrd and 6-mercaptoguanosine.
机译:在分离自人神经母细胞瘤的克隆细胞系IMR-32的膜制剂中,检测到与血型相关糖鞘脂的体外生物合成有关的两种不同的糖脂:岩藻糖基转移酶活性。该膜制剂包含α(1,2)-岩藻糖基转移酶(EC 2.4.1.89),其催化将蔗糖从GDP-[14C]岩藻糖转移至新内酯四糖基神经酰胺或新内酯五糖基神经酰胺,分别形成H-1型和B-1型糖脂。第二个岩藻糖基转移酶催化岩藻糖转移至乳三糖基神经酰胺[GlcNAc(β1-3)Gal(β1-41-4)Glc-Cer],形成新型Lea型糖脂的四糖基神经酰胺中间体。在类似条件下测试时,UDP-半乳糖:乳糖三酰基神经酰胺β-半乳糖基转移酶(EC 2.4.1.86)的活性是UDP-半乳糖:α-半乳糖基转移酶(EC 2.4.1.87)的4倍。还比较了在存在4微米BrdUrd和6-巯基鸟苷的情况下分化的细胞中的α-岩藻糖基转移酶活性以及[14C]岩藻糖掺入糖蛋白和糖脂的能力。

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