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 Lee 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 (US)
Pages (from-to)482-489
Number of pages8
JournalCarbohydrate Research
Volume342
Issue number3-4
DOIs
StatePublished - Feb 26 2007

Fingerprint

G(M1) Ganglioside
Metabolomics
Lasers
Fluorescence
Galactosidases
Sialyltransferases
Acylation
Capillary electrophoresis
Glycolipids
Capillary Electrophoresis
Neuraminidase
Alanine
Esters
Derivatives
Molecules
Enzymes
tetramethylrhodamine
ganglio-N-triaosylceramide

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Organic Chemistry

Cite this

Synthesis of reference standards to enable single cell metabolomic studies of tetramethylrhodamine-labeled ganglioside GM1. / Larsson, E. Andreas; Olsson, Ulf; Whitmore, Colin D.; Martins, Rita; Tettamanti, Guido; Schnaar, Ronald Lee; Dovichi, Norman J.; Palcic, Monica M.; Hindsgaul, Ole.

In: Carbohydrate Research, Vol. 342, No. 3-4, 26.02.2007, p. 482-489.

Research output: Contribution to journalArticle

Larsson, EA, Olsson, U, Whitmore, CD, Martins, R, Tettamanti, G, Schnaar, RL, Dovichi, NJ, Palcic, MM & Hindsgaul, O 2007, 'Synthesis of reference standards to enable single cell metabolomic studies of tetramethylrhodamine-labeled ganglioside GM1', Carbohydrate Research, vol. 342, no. 3-4, pp. 482-489. https://doi.org/10.1016/j.carres.2006.10.002
Larsson, E. Andreas ; Olsson, Ulf ; Whitmore, Colin D. ; Martins, Rita ; Tettamanti, Guido ; Schnaar, Ronald Lee ; Dovichi, Norman J. ; Palcic, Monica M. ; Hindsgaul, Ole. / Synthesis of reference standards to enable single cell metabolomic studies of tetramethylrhodamine-labeled ganglioside GM1. In: Carbohydrate Research. 2007 ; Vol. 342, No. 3-4. pp. 482-489.
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