Apolipoprotein A-I modulates HDL particle size in the absence of apolipoprotein A-II
| Autor: | Rachel Hart, Carissa Thornock, Shimpi Bedi, Zsuzsanna Kuklenyik, John T. Melchior, Jay W. Heinecke, Tomas Vaisar, Scott E. Street, Jere P. Segrest, Amy S. Shah, W. Sean Davidson, Noemie Clouet-Foraison, Jay Jerome |
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| Rok vydání: | 2021 |
| Předmět: |
SEC
size-exclusion chromatography Apolipoprotein B Size-exclusion chromatography Apolipoprotein A-II APOA2 apolipoprotein A-II POPC palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine QD415-436 high-density lipoprotein UC-HDL HDL isolated by ultracentrifugation Biochemistry BS3 bis-(sulfosuccinimidyl) suberate PC phosphatidylcholine chemistry.chemical_compound HDL subfractions FPLC fast protein liquid chromatography Endocrinology High-density lipoprotein Affinity chromatography LpA-I lipoproteins with APOA1 and no APOA2 Cholesterylester transfer protein CETP cholesteryl ester transfer protein Particle Size mass spectrometry GO gene ontology Apolipoprotein A-I biology Chemistry Cholesterol HDL APOA1 apolipoprotein A-I isotope-dilution MS/MS differential ion mobility analysis Fast protein liquid chromatography Cell Biology LpA-IS small subfraction of LpA-I ratchet model LpA-IL large subfraction of LpA-I lipoproteins size-exclusion chromatography IAC immunoaffinity chromatography composition biology.protein Biophysics Particle lipids (amino acids peptides and proteins) Research Article LpA-I/A-II lipoproteins with APOA1 and APOA2 |
| Zdroj: | Journal of Lipid Research Journal of Lipid Research, Vol 62, Iss, Pp 100099-(2021) |
| ISSN: | 0022-2275 |
| DOI: | 10.1016/j.jlr.2021.100099 |
| Popis: | Human high-density lipoproteins (HDLs) are a complex mixture of structurally related nanoparticles that perform distinct physiological functions. We previously showed that human HDL containing apolipoprotein A-I (APOA1) but not apolipoprotein A-II (APOA2), designated LpA-I, is composed primarily of two discretely sized populations. Here, we isolated these particles directly from human plasma by antibody affinity chromatography, separated them by high-resolution size-exclusion chromatography and performed a deep molecular characterization of each species. The large and small LpA-I populations were spherical with mean diameters of 109 A and 91 A, respectively. Unexpectedly, isotope dilution MS/MS with [15N]-APOA1 in concert with quantitation of particle concentration by calibrated ion mobility analysis demonstrated that the large particles contained fewer APOA1 molecules than the small particles; the stoichiometries were 3.0 and 3.7 molecules of APOA1 per particle, respectively. MS/MS experiments showed that the protein cargo of large LpA-I particles was more diverse. Human HDL and isolated particles containing both APOA1 and APOA2 exhibit a much wider range and variation of particle sizes than LpA-I, indicating that APOA2 is likely the major contributor to HDL size heterogeneity. We propose a ratchet model based on the trefoil structure of APOA1 whereby the helical cage maintaining particle structure has two “settings”—large and small—that accounts for these findings. This understanding of the determinants of HDL particle size and protein cargo distribution serves as a basis for determining the roles of HDL subpopulations in metabolism and disease states. |
| Databáze: | OpenAIRE |
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