title: Glycomic analysis of high density lipoprotein shows a highly sialylated particle.
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 (Hexose5HexNAc4Neu5Ac2) 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.
authors.affiliation: Department of Chemistry, ‡Department of Food Science and Technology, and §Foods for Health Institute, University of California , Davis, California 95616, United States.
authors.forename: Jincui
authors.initials: J
authors.lastname: Huang
authors.forename: Hyeyoung
authors.initials: H
authors.lastname: Lee
authors.forename: Angela M
authors.initials: AM
authors.lastname: Zivkovic
authors.forename: Jennifer T
authors.initials: JT
authors.lastname: Smilowitz
authors.forename: Nancy
authors.initials: N
authors.lastname: Rivera
authors.forename: J Bruce
authors.initials: JB
authors.lastname: German
authors.forename: Carlito B
authors.initials: CB
authors.lastname: Lebrilla
mesh_terms: D000595:Amino Acid Sequence; D002241:Carbohydrates / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D005732:Gangliosides / Q000737:chemistry; D008075:Lipoproteins, HDL / Q000737:chemistry*; D008969:Molecular Sequence Data; D019158:N-Acetylneuraminic Acid / Q000737:chemistry*; D053719:Tandem Mass Spectrometry
chemical_list: D002241:Carbohydrates; D005732:Gangliosides; D008075:Lipoproteins, HDL; D019158:N-Acetylneuraminic Acid