Synthesis of reference standards to enable single cell metabolomic studies of tetramethylrhodamine-labeled ganglioside GM1

E. Andreas Larsson, Ulf Olsson, Colin D. Whitmore, Rita Martins, Guido Tettamanti, Ronald L. Schnaar, Norman J. Dovichi, Monica M. Palcic, Ole Hindsgaul

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Pages (from-to)482-489
Number of pages8
JournalCarbohydrate Research
Volume342
Issue number3-4
DOIs
Publication statusPublished - Feb 26 2007

Keywords

  • Fluorescence labeling
  • Ganglioside
  • Glycolipid metabolism
  • GM1
  • Membrane recycling
  • Metabolomics

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Organic Chemistry

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    Larsson, E. A., Olsson, U., Whitmore, C. D., Martins, R., Tettamanti, G., Schnaar, R. L., Dovichi, N. J., Palcic, M. M., & Hindsgaul, O. (2007). Synthesis of reference standards to enable single cell metabolomic studies of tetramethylrhodamine-labeled ganglioside GM1. Carbohydrate Research, 342(3-4), 482-489. https://doi.org/10.1016/j.carres.2006.10.002