Synthesis of reference standards to enable single cell metabolomic studies of tetramethylrhodamine-labeled ganglioside GM1
- 作者：E. Andreas Larsson ; Ulf Olsson ; Colin D. Whitmore ; Rita Martins ; Guido Tettamanti ; Ronald L. Schnaar ; Norman J. Dovichi ; Monica M. Palcic ; Ole Hindsgaul
- 关键词：Glycolipid metabolism ; Ganglioside ; GM1 ; Metabolomics ; Fluorescence labeling ; Membrane recycling
- 刊名：Carbohydrate Research
- 出版年：2007
- 期刊代码：15_00086215
- 类别：ch
- 出版时间：26 February 2007
- 卷：342
- 期：3-4
- 页码：482-489
- 文件大小：349 K
摘要
Ganglioside GM1 and its seven potential catabolic products: asialo-GM1, GM2, asialo-GM2, GM3, Lac-Cer, Glc-Cer and Cer, were labeled with tetramethylrhodamine (TMR) to permit ultra-sensitive analysis using laser-induced fluorescence (LIF) detection. The preparation involved acylation of the homogenous C18 lyso-forms of GM1, Lac-Cer, Glc-Cer and Cer with the N-hydroxysuccinimide ester of a β-alanine-tethered 6-TMR derivative, followed by conversion of these labeled products using galactosidase, sialidase, and sialyltransferase enzymes. The TMR–glycolipid analogs produced are detectable on TLC down to the 1 ng level by the naked eye. All eight compounds could be separated within 4 min in capillary electrophoresis where they could be detected at the zeptomole (ca. 1000 molecule) level using LIF.