Monosialogangliosides of human myelogenous leukemia HL60 cells and normal human leukocytes. 1. Separation of E-selectin binding from nonbinding gangliosides, and absence of sialosyl-Le(x) having tetraosyl to octaosyl core.
Monosialogangliosides of human myelogenous leukemia HL60 cells and normal human leukocytes. 1. Separation of E-selectin binding from nonbinding gangliosides, and absence of sialosyl-Le(x) having tetraosyl to octaosyl core.
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人骨髓性白血病 HL60 细胞和正常人白细胞的单唾液酸神经节苷脂。
DOI：
10.1021/bi951600r
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发表时间：
1996
期刊：
影响因子：
2.9
通讯作者：
Reinhold,WN
中科院分区：
文献类型：
--
作者：
Stroud,MR;Handa,K;Salyan,ME;Ito,K;Levery,SB;Hakomori,S;Reinhold,BB;Reinhold,WN
以往的研究表明，唾液酸脂(sialosyl-lex，SLeX)是一种在人中性粒细胞和髓系白血病HL60细胞中表达的配体，与E-选择素和可能的P-选择素结合。然而，缺乏关于这些细胞中碳水化合物表位结构的明确数据。因此，一项系统的研究开始了，使用了大量的HL-60细胞(包装的≥为1200毫升)和人的白细胞(包装的≈为100毫升)。提取神经节苷脂，然后广泛分级，并检查每个组分的E-和P-选择素结合能力。 (I)只有以~gt；10个单糖单元(或~gt；4N-乙酰乳糖胺单元)为核心的单唾液酸神经节苷脂在静态条件下与E-选择素结合，用32P标记的表达E-选择素的CHO细胞的薄层层析覆盖技术。(Ii)在结合组分中没有检测到硫酸盐基团，在这些条件下，二唾液酸神经节苷脂和三唾液酸神经节苷组分不显示E-选择素结合。(Iii)在使用32P标记的表达P-选择素的CHO细胞的类似检测系统下，没有一个组分显示P-选择素结合。(4)HL60细胞的主要神经节苷脂为结构I−XI(如正文表1所示)，在上述条件下均未显示E-选择素结合。(V)上皮性肿瘤的主要神经节苷脂(如表2所示)中，HL60细胞和中性粒细胞中完全不存在具有四糖至八糖神经酰胺核心的SLEX神经节苷脂。本文报道了神经节苷脂I−XI的分离和化学性质。
Previous studies suggested that sialosyl-Lex(SLex) is a ligand expressed in human neutrophils and myelogenous leukemia HL60 cells which binds to E-selectin and possibly P-selectin. However, clear data on structures of carbohydrate epitopes in these cells were lacking. A systematic study was therefore initiated, employing a large quantity of HL60 cells (≥1200 mL packed) and human leukocytes (≈100 mL packed). Gangliosides were extracted, followed by extensive fractionation and examination of the E- and P-selectin binding ability of each fraction. The following results were of particular interest: (i) Only monosialogangliosides having a polylactosamine core with >10 monosaccharide units (or >4N-acetyllactosamine units) showed E-selectin binding under static conditions with thin-layer chromatography overlay technique employing32P-labeled E-selectin-expressing CHO cells. (ii) Sulfate groups were not detectable in the binding fractions, and di- and trisialoganglioside fractions did not show E-selectin binding under these conditions. (iii) None of the fractions showed P-selectin binding under a similar assay system using32P-labeled P-selectin-expressing CHO cells. (iv) Major gangliosides of HL60 cells were structuresI−XI(shown in Table 1 of text), none of which showed E-selectin binding under the above conditions. (v) SLexgangliosides having tetraosyl to octaosyl ceramide core, which are the major gangliosides of epithelial tumors (shown in Table 2), were completely absent from HL60 cells and neutrophils. Isolation and chemical characterization of ganglioside structuresI−XIare described in this paper.