Glycomic analysis of high density lipoprotein shows a highly sialylated particle

Jincui Huang, Hyeyoung Lee, Angela M. Zivkovic, Jennifer T. Smilowitz, Nancy Rivera, J. Bruce German, Carlito B Lebrilla

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. In this study, HDL was isolated from plasma by sequential micro-ultracentrifugation, followed by glycoprotein and glycolipid analysis. N-Glycans, glycopeptides, and gangliosides were extracted and purified followed by analysis with nano-HPLC Chip quadrupole time of flight mass spectrometry and MS/MS. HDL particles were found to be highly sialylated. Most of the N-glycans (∼90%) from HDL glycoproteins were sialylated with one or two neuraminic acids (Neu5Ac). The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc 4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. A 60% GM3 and 40% GD3 distribution was observed. Both GM3 and GD3 were composed of heterogeneous ceramide lipid tails, including d18:1/16:0 and d18:1/23:0. This report describes for the first time a glycomic approach for analyzing HDL, highlighting that HDL are highly sialylated particles.

Original languageEnglish (US)
Pages (from-to)681-691
Number of pages11
JournalJournal of Proteome Research
Volume13
Issue number2
DOIs
StatePublished - Feb 7 2014

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Glycomics
HDL Lipoproteins
Polysaccharides
Glycoproteins
Gangliosides
alpha-2-HS-Glycoprotein
Neuraminic Acids
Apolipoprotein C-III
Sialic Acids
Glycosylation
Lipids
Glycoconjugates
Glycopeptides
Ceramides
Ultracentrifugation
Glycolipids
Mass spectrometry
Mass Spectrometry
High Pressure Liquid Chromatography
Plasmas

Keywords

  • gangliosides
  • glycomics
  • glycoproteomics
  • HDL
  • mass spectrometry
  • sialylation

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

Cite this

Huang, J., Lee, H., Zivkovic, A. M., Smilowitz, J. T., Rivera, N., German, J. B., & Lebrilla, C. B. (2014). Glycomic analysis of high density lipoprotein shows a highly sialylated particle. Journal of Proteome Research, 13(2), 681-691. https://doi.org/10.1021/pr4012393

Glycomic analysis of high density lipoprotein shows a highly sialylated particle. / Huang, Jincui; Lee, Hyeyoung; Zivkovic, Angela M.; Smilowitz, Jennifer T.; Rivera, Nancy; German, J. Bruce; Lebrilla, Carlito B.

In: Journal of Proteome Research, Vol. 13, No. 2, 07.02.2014, p. 681-691.

Research output: Contribution to journalArticle

Huang, J, Lee, H, Zivkovic, AM, Smilowitz, JT, Rivera, N, German, JB & Lebrilla, CB 2014, 'Glycomic analysis of high density lipoprotein shows a highly sialylated particle', Journal of Proteome Research, vol. 13, no. 2, pp. 681-691. https://doi.org/10.1021/pr4012393
Huang J, Lee H, Zivkovic AM, Smilowitz JT, Rivera N, German JB et al. Glycomic analysis of high density lipoprotein shows a highly sialylated particle. Journal of Proteome Research. 2014 Feb 7;13(2):681-691. https://doi.org/10.1021/pr4012393
Huang, Jincui ; Lee, Hyeyoung ; Zivkovic, Angela M. ; Smilowitz, Jennifer T. ; Rivera, Nancy ; German, J. Bruce ; Lebrilla, Carlito B. / Glycomic analysis of high density lipoprotein shows a highly sialylated particle. In: Journal of Proteome Research. 2014 ; Vol. 13, No. 2. pp. 681-691.
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abstract = "Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. In this study, HDL was isolated from plasma by sequential micro-ultracentrifugation, followed by glycoprotein and glycolipid analysis. N-Glycans, glycopeptides, and gangliosides were extracted and purified followed by analysis with nano-HPLC Chip quadrupole time of flight mass spectrometry and MS/MS. HDL particles were found to be highly sialylated. Most of the N-glycans (∼90{\%}) from HDL glycoproteins were sialylated with one or two neuraminic acids (Neu5Ac). The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc 4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. A 60{\%} GM3 and 40{\%} GD3 distribution was observed. Both GM3 and GD3 were composed of heterogeneous ceramide lipid tails, including d18:1/16:0 and d18:1/23:0. This report describes for the first time a glycomic approach for analyzing HDL, highlighting that HDL are highly sialylated particles.",
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