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Analysis of ubiquinones, dolichols, and dolichol diphosphate-oligosaccharides by liquid chromatography-electrospray ionization-mass spectrometry.

Publication ,  Journal Article
Garrett, TA; Guan, Z; Raetz, CRH
Published in: Methods Enzymol
2007

Prenols, a class of lipids formed by the condensation of five carbon isoprenoids, have important roles in numerous metabolic pathways of the eukaryotic cell. Prenols are found in the cell as free alcohols, such as dolichol, or can be attached to vitamins, as with the fat soluble vitamins. In addition, prenols such as farnesyl- and geranylgeranyl-diphosphate are substrates for the transfer of farnesyl and geranylgeranyl units to proteins with important implications for signal transduction within the cell. Dolichol phosphate- and dolichol diphosphate-linked sugars are central to the formation of the lipid-linked branched oligosaccharide, Dol-PP-(GlcNAc)2(Man)9(Glc)3, used for co-translational en bloc protein N-glycosylation in the lumen of the endoplasmic reticulum. Toward furthering our understanding of the role of prenol lipids in the cell, we have developed a method for the detection and quantification of dolichol and coenzyme Q by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS). These methods, developed using the mouse macrophage RAW 264.7 tumor cells, are broadly applicable to other cell lines, tissues, bacteria, and yeast. We also present a new MS-based method for the detection and structural characterization of the intact dolichol diphosphate oligosaccharide Dol-PP-(GlcNAc)2 (Man)9(Glc)3 from porcine pancreas.

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Published In

Methods Enzymol

DOI

ISSN

0076-6879

Publication Date

2007

Volume

432

Start / End Page

117 / 143

Location

United States

Related Subject Headings

  • Ubiquinone
  • Spectrometry, Mass, Electrospray Ionization
  • Oligosaccharides
  • Dolichols
  • Chromatography, Liquid
  • Biochemistry & Molecular Biology
  • 3101 Biochemistry and cell biology
  • 0601 Biochemistry and Cell Biology
 

Citation

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Garrett, T. A., Guan, Z., & Raetz, C. R. H. (2007). Analysis of ubiquinones, dolichols, and dolichol diphosphate-oligosaccharides by liquid chromatography-electrospray ionization-mass spectrometry. Methods Enzymol, 432, 117–143. https://doi.org/10.1016/S0076-6879(07)32005-3
Garrett, Teresa A., Ziqiang Guan, and Christian R. H. Raetz. “Analysis of ubiquinones, dolichols, and dolichol diphosphate-oligosaccharides by liquid chromatography-electrospray ionization-mass spectrometry.Methods Enzymol 432 (2007): 117–43. https://doi.org/10.1016/S0076-6879(07)32005-3.
Garrett, Teresa A., et al. “Analysis of ubiquinones, dolichols, and dolichol diphosphate-oligosaccharides by liquid chromatography-electrospray ionization-mass spectrometry.Methods Enzymol, vol. 432, 2007, pp. 117–43. Pubmed, doi:10.1016/S0076-6879(07)32005-3.
Journal cover image

Published In

Methods Enzymol

DOI

ISSN

0076-6879

Publication Date

2007

Volume

432

Start / End Page

117 / 143

Location

United States

Related Subject Headings

  • Ubiquinone
  • Spectrometry, Mass, Electrospray Ionization
  • Oligosaccharides
  • Dolichols
  • Chromatography, Liquid
  • Biochemistry & Molecular Biology
  • 3101 Biochemistry and cell biology
  • 0601 Biochemistry and Cell Biology