title: Biochemical and functional characterization of charge-defined subfractions of high-density lipoprotein from normal adults.
abstract: High-density lipoprotein (HDL) is regarded as atheroprotective because it provides antioxidant and anti-inflammatory benefits and plays an important role in reverse cholesterol transport. In this paper, we outline a novel methodology for studying the heterogeneity of HDL. Using anion-exchange chromatography, we separated HDL from 6 healthy individuals into five subfractions (H1 through H5) with increasing charge and evaluated the composition and biologic activities of each subfraction. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis showed that apolipoprotein (apo) AI and apoAII were present in all 5 subfractions; apoCI was present only in H1, and apoCIII and apoE were most abundantly present in H4 and H5. HDL-associated antioxidant enzymes such as lecithin-cholesterol acyltransferase, lipoprotein-associated phospholipase A2, and paraoxonase 1 were most abundant in H4 and H5. Lipoprotein isoforms were analyzed in each subfraction by using matrix-assisted laser desorption-time-of-flight mass spectrometry. To quantify other proteins in the HDL subfractions, we used the isobaric tags for the relative and absolute quantitation approach followed by nanoflow liquid chromatography-tandem mass spectrometry analysis. Most antioxidant proteins detected were found in H4 and H5. The ability of each subfraction to induce cholesterol efflux from macrophages increased with increasing HDL electronegativity, with the exception of H5, which promoted the least efflux activity. In conclusion, anion-exchange chromatography is an attractive method for separating HDL into subfractions with distinct lipoprotein compositions and biologic activities. By comparing the properties of these subfractions, it may be possible to uncover HDL-specific proteins that play a role in disease.
authors.affiliation: L5 Research Center, Medical Research Department, China Medical University Hospital, 2 Yude Road, North District, Taichung, 40447, Taiwan.
authors.forename: Ju-Yi
authors.initials: JY
authors.lastname: Hsieh
authors.affiliation: L5 Research Center, Medical Research Department, China Medical University Hospital, 2 Yude Road, North District, Taichung, 40447, Taiwan.|College of Medicine, China Medical University, Taichung, 40402, Taiwan.
authors.forename: Chiz-Tzung
authors.initials: CT
authors.lastname: Chang
authors.affiliation: Section of Atherosclerosis and Lipoprotein Research, Department of Medicine, Baylor College of Medicine, Houston, Texas 77030, United States.
authors.forename: Max T
authors.initials: MT
authors.lastname: Huang
authors.affiliation: L5 Research Center, Medical Research Department, China Medical University Hospital, 2 Yude Road, North District, Taichung, 40447, Taiwan.
authors.forename: Chia-Ming
authors.initials: CM
authors.lastname: Chang
authors.affiliation: L5 Research Center, Medical Research Department, China Medical University Hospital, 2 Yude Road, North District, Taichung, 40447, Taiwan.
authors.forename: Chia-Ying
authors.initials: CY
authors.lastname: Chen
authors.affiliation: L5 Research Center, Medical Research Department, China Medical University Hospital, 2 Yude Road, North District, Taichung, 40447, Taiwan.|Graduate Institute of Clinical Medical Science, China Medical University, Taichung, 40402, Taiwan.
authors.forename: Ming-Yi
authors.initials: MY
authors.lastname: Shen
authors.affiliation: Proteomics Core Laboratory, Department of Medical Research, China Medical University Hospital, Taichung, 40402, Taiwan.
authors.forename: Hsin-Yi
authors.initials: HY
authors.lastname: Liao
authors.affiliation: Graduate Institute of Clinical Medical Science, China Medical University, Taichung, 40402, Taiwan.|Department of Medical Research, China Medical University Hospital, Taichung, 40402, Taiwan.|Department of Health and Nutrition Biotechnology, Asia University, Taichung 41354, Taiwan.
authors.forename: Guei-Jane
authors.initials: GJ
authors.lastname: Wang
authors.affiliation: L5 Research Center, Medical Research Department, China Medical University Hospital, 2 Yude Road, North District, Taichung, 40447, Taiwan.|Graduate Institute of Clinical Medical Science, China Medical University, Taichung, 40402, Taiwan.|Section of Atherosclerosis and Lipoprotein Research, Department of Medicine, Baylor College of Medicine, Houston, Texas 77030, United States.|Vascular and Medicinal Research, Texas Heart Institute, Houston, Texas 77030, United States.
authors.forename: Chu-Huang
authors.initials: CH
authors.lastname: Chen
authors.affiliation: Proteomics Core Laboratory, Department of Medical Research, China Medical University Hospital, Taichung, 40402, Taiwan.|Section of Atherosclerosis and Lipoprotein Research, Department of Medicine, Baylor College of Medicine, Houston, Texas 77030, United States.
authors.forename: Chao-Jung
authors.initials: CJ
authors.lastname: Chen
authors.affiliation: L5 Research Center, Medical Research Department, China Medical University Hospital, 2 Yude Road, North District, Taichung, 40447, Taiwan.|Graduate Institute of Clinical Medical Science, China Medical University, Taichung, 40402, Taiwan.|Section of Atherosclerosis and Lipoprotein Research, Department of Medicine, Baylor College of Medicine, Houston, Texas 77030, United States.|Vascular and Medicinal Research, Texas Heart Institute, Houston, Texas 77030, United States.
authors.forename: Chao-Yuh
authors.initials: CY
authors.lastname: Yang
mesh_terms: D000328:Adult; D000837:Anion Exchange Resins / Q000737:chemistry; D005591:Chemical Fractionation / Q000379:methods*; D005260:Female; D006801:Humans; D008075:Lipoproteins, HDL / Q000032:analysis* / Q000737:chemistry*; D008297:Male; D008875:Middle Aged
chemical_list: D000837:Anion Exchange Resins; D008075:Lipoproteins, HDL