GLYCOSPHINGOLIPIDS FROM BOVINE RED BLOOD CELLS & FOETAL CALF BRAIN

来自牛红细胞的糖鞘脂

• 批准号: 6123271
• 负责人: ROBERT H MCCLUER
• 金额: --
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-01 至 1999-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6123271
• 关键词: biological products; biomaterials; biomedical resource; carbohydrates; growth /development; hematology; lipids; nervous system; spectrometry; structural biology; technology /technique

Stage-specific embryonic antigen (SSEA) glycosphingolipids (GSLs) found in the central nervous system are impficated in regulating cell-cell recognition, targeting and migration of cells during development. Through the action of fucosyltransferase enzymes, SSEA-1 (Lewis~ glycolipids arebiosynthesized in the brain by fucosylation of lipid substrates with the neolacto series glycolipid core structure [Galp 1-4GIcNAcp 1-3Galp 1-4GIcp I- I'Cer] (originally termed paragloboside) or its higher analogs. In order to optimize methodology for high sensitivity structural determinations of SSEA-1 type glycofipids from fetal calf brain, potential precursors and SSEA-1 glycolipids of previously established structure were first isolated from bovine erythrocytes and beige mutant mouse kidney, purified by colurnn chromatography and characterized by MALDI-TOF MS, LSIM[S, and M[S/M[S, among other techniques. Peracetylated derivatives were detected at the low ferntomole level by MALDI-TOF MS and the subnanomole level by LSIMS. MALDI-TOF MS produced mainly [M+Na]' and [M+K]' species. On the basis of the direct and tandem mass spectral analyses of peracetylated and permethylated derivatives, the carbohydrate sequences in the selected bovine erythrocyte and mouse kidney GSL fractions were found to be consistent with those of glycolipids previously reported from larger-scale studies of these sources. Their heterogeneous ceramide moieties were characterized by CID M[S/M[S of abundant Zo-type ions in the LS1 mass spectra of the permethylated GSLs. MALDI-PSD-TOF mass spectral analyses of low and subpicomole amounts of derivatized GSL fractions from fetal calf brain provided carbohydrate sequence information that indicates the presence of mono- and difucosylated SSEA-I neolacto series glycolipids and their nonfucosylated analogs.

阶段特异性的胚胎抗原（SSEA）糖磷脂（GSLS） 在中枢神经系统中发现在调节中受到阻碍 细胞细胞识别，靶向和迁移细胞在 发展。 通过岩藻糖基转移酶的作用，SSEA-1 （刘易斯〜糖脂是通过构造的 脂质底物具有新乳清系列糖脂核心结构 [GALP 1-4GICNACP 1-3GALP 1-4GICP I- I'CER]（最初称为 paragloboside）或其较高的类似物。 为了优化 SSEA-1高灵敏度结构测定的方法 从胎牛大脑，潜在的前体和 先前建立的结构的SSEA-1糖脂首先 从牛红细胞和米色突变小鼠肾脏中分离出来， 由Colurnn色谱纯化，以MALDI-TOF MS为特征 lsim [s和m [s/m [s，以及其他技术。 过乙酰化 MALDI-TOF MS在低蕨类酚水平上检测到衍生物 以及LSIMS的亚纳米尔水平。 MALDI-TOF MS主要产生 [m+na]和[m+k]'物种。 基于直接和串联 过乙酰化和苄苄苄化衍生物的质谱分析， 选定的牛红细胞中的碳水化合物序列和 发现鼠标肾脏GSL分数与 糖脂先前从对这些的大规模研究中报道 来源。 他们的异质神经酰胺部分的特征是 cid m [s/m [s/m [s的ls1质量光谱中丰富的ZO型离子 苄氯化GSL。 低和低的MALDI-PSD-TOF质谱分析 从胎牛大脑的衍生化GSL级分的亚匹素量 提供的碳水化合物序列信息表明存在 单核和沉核酸的SSEA-I NEOLACTO系列糖脂和 它们的非二核类似类似物。