制备溶血神经节苷脂GM1和溶血神经节苷脂GM2的简便方法。

Facile method for the preparation of lyso-GM1 and lyso-GM2.

作者信息

Ando Takayuki, Li Su-Chen, Ito Makoto, Li Yu-Teh

机构信息

Department of Biochemistry, Tulane University Health Sciences Center School of Medicine, New Orleans, LA 70112, USA.

出版信息

J Chromatogr A. 2005 Jun 17;1078(1-2):193-5. doi: 10.1016/j.chroma.2005.04.058.

DOI:10.1016/j.chroma.2005.04.058

PMID:16010718

Abstract

This paper reports a facile method for the preparation of lyso-GM1 [Gal beta1-->3GalNAc beta1--> 4(Neu5Ac alpha2-->3)Galbeta1-->4Glc beta1-->1'-sphingosine] and lyso-GM2 [GalNAc beta1-->4(Neu5Ac alpha2-->3)Gal beta1-->4Glc beta1-->sphingosine], respectively, from GM1 [Galbeta1-->3GalNAc beta1-->4(Neu5Ac alpha2-->3)Galbeta1-->4Glc beta1-->1'-Cer] and GM2[GalNAc beta1-->4(Neu5Ac alpha2-->3)Galbeta1-->4Glc beta1-->1'-Cer], using sphingolipid ceramide deacylase and high performance anion-exchange chromatography (HPAEC). The enzymatically released lyso-GM1 and/or lyso-GM2 was effectively separated from its parent ganglioside by HPAEC using a Mono Q HR 5/5 column with an Amersham Biosciences fast protein liquid chromatography system. The yield was almost quantitative and the separation completed in approximately 3 h. This method is more convenient and effective than the conventional method using alkaline hydrolysis and silicic acid chromatography to generate and purify lyso-gangliosides.

摘要

本文报道了一种简便的方法，分别从GM1[Galβ1→3GalNAcβ1→4(Neu5Acα2→3)Galβ1→4Glcβ1→1'-神经酰胺]和GM2[GalNAcβ1→4(Neu5Acα2→3)Galβ1→4Glcβ1→1'-神经酰胺]制备溶血GM1[Galβ1→3GalNAcβ1→4(Neu5Acα2→3)Galβ1→4Glcβ1→鞘氨醇]和溶血GM2[GalNAcβ1→4(Neu5Acα2→3)Galβ1→4Glcβ1→鞘氨醇]，采用鞘脂神经酰胺脱酰酶和高效阴离子交换色谱法（HPAEC）。使用带有Amersham Biosciences快速蛋白质液相色谱系统的Mono Q HR 5/5柱，通过HPAEC将酶促释放的溶血GM1和/或溶血GM2与其母体神经节苷脂有效分离。产率几乎是定量的，分离在大约3小时内完成。该方法比使用碱性水解和硅酸色谱法生成和纯化溶血神经节苷脂的传统方法更方便、有效。