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神经节苷脂GD1 alpha' IV3Neu5Ac，III6Neu5Ac-GgOse4Cer，是高转移性小鼠淋巴网状肿瘤细胞系MDAY-D2中的一种主要的双唾液酸神经节苷脂。

The ganglioside GD1 alpha' IV3Neu5Ac, III6Neu5Ac-GgOse4Cer, is a major disialoganglioside in the highly metastatic murine lymphoreticular tumour cell line MDAY-D2.

作者信息

Müthing J, Peter-Katalinić J, Hanisch F G, Unland F, Lehmann J

机构信息

Institut für Zellkulturtechnik der Universität, Bielefeld, Germany.

出版信息

Glycoconj J. 1994 Apr;11(2):153-62. doi: 10.1007/BF00731155.

DOI:10.1007/BF00731155
PMID:7804005
Abstract

The aim of the present study was to investigate the ganglioside expression of the highly metastatic murine lymphoreticular tumour cell line MDAY-D2. Cells were propagated under controlled pH conditions and oxygen supply in bioreactors of 1 and 7.5 l volumes by repeated batch fermentation. Gangliosides were isolated from 2.7 x 10(11) cells, purified by silica gel chromatography and separated into mono- and disialoganglioside fractions by preparative DEAE anion exchange high performance liquid chromatography. Individual gangliosides were obtained by preparative thin layer chromatography. Their structural features were established by immunostaining, fast atom bombardment and gas chromatography mass spectrometry. In addition to gangliosides of the GM1a-pathway (GM2, GM1a and GD1a) and GM1b (IV3Neu5Ac-GgOse4Cer) and GalNAc-GM1b of the Gm1b-pathway, the disialoganglioside GD1 alpha (IV3Neu5Ac, III6Neu5Ac-GgOse4Cer) was found in equal amounts compared to GD1a (IV3Neu5Ac, II3Neu5Ac-GgOse4Cer). All gangliosides were substituted with C24:0, 24:1 and C16:0 fatty acids, sphingosine and N-acetylneuraminic acid as the sole sialic acid.

摘要

本研究的目的是调查高转移性小鼠淋巴网状肿瘤细胞系MDAY-D2的神经节苷脂表达情况。通过重复分批发酵，在1升和7.5升体积的生物反应器中，在受控的pH条件和氧气供应下培养细胞。从2.7×10¹¹个细胞中分离出神经节苷脂，通过硅胶柱色谱法纯化，并通过制备性DEAE阴离子交换高效液相色谱法分离成单唾液酸和双唾液酸神经节苷脂馏分。通过制备性薄层色谱法获得单个神经节苷脂。通过免疫染色、快原子轰击和气相色谱-质谱法确定其结构特征。除了GM1a途径的神经节苷脂（GM2、GM1a和GD1a）以及GM1b途径的GM1b（IV3Neu5Ac-GgOse4Cer）和GalNAc-GM1b外，还发现双唾液酸神经节苷脂GD1α（IV3Neu5Ac，III6Neu5Ac-GgOse4Cer）与GD1a（IV3Neu5Ac，II3Neu5Ac-GgOse4Cer）含量相等。所有神经节苷脂均被C24:0、24:1和C16:0脂肪酸、鞘氨醇和N-乙酰神经氨酸（作为唯一的唾液酸）取代。

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