Mass spectrometry‐based analytical tools for the molecular protein characterization of human plasma lipoproteins
- 28 June 2005
- journal article
- research article
- Published by Wiley in Proteomics
- Vol. 5 (10), 2619-2630
- https://doi.org/10.1002/pmic.200401233
Abstract
Lipoproteins are a heterogeneous population of blood plasma particles composed of apolipoproteins and lipids. Lipoproteins transport exogenous and endogenous triglycerides and cholesterol from sites of absorption and formation to sites of storage and usage. Three major classes of lipoproteins are distinguished according to their density: high‐density (HDL), low‐density (LDL) and very low‐density lipoproteins (VLDL). While HDLs contain mainly apolipoproteins of lower molecular weight, the two other classes contain apolipoprotein B and apolipoprotein (a) together with triglycerides and cholesterol. HDL concentrations were found to be inversely related to coronary heart disease and LDL/VLDL concentrations directly related. Although many studies have been published in this area, few have concentrated on the exact protein composition of lipoprotein particles. Lipoproteins were separated by density gradient ultracentrifugation into different subclasses. Native gel electrophoresis revealed different gel migration behaviour of the particles, with less dense particles having higher apparent hydrodynamic radii than denser particles. Apolipoprotein composition profiles were measured by matrix‐assisted laser desorption/ionization‐mass spectrometry on a macromizer™ instrument, equipped with the recently introduced cryodetector technology, and revealed differences in apolipoprotein composition between HDL subclasses. By combining these profiles with protein identifications from native and denaturing polyacrylamide gels by liquid chromatography‐tandem mass spectrometry, we characterized comprehensively the exact protein composition of different lipoprotein particles. We concluded that the differential display of protein weight information acquired by macromizer™ mass spectrometry is an excellent tool for revealing structural variations of different lipoprotein particles, and hence the foundation is laid for the screening of cardiovascular disease risk factors associated with lipoproteins.Keywords
This publication has 26 references indexed in Scilit:
- An investigation into the human serum “interactome”Electrophoresis, 2004
- Clinical Proteomics: Are We There Yet?Analytical Chemistry, 2003
- Analysis of High-Density Lipoprotein Apolipoproteins Recovered from Specific Immobilized pH Gradient Gel pI Domains by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass SpectrometryAnalytical Chemistry, 2003
- Assembly and Secretion of Very Low Density Lipoproteins Containing Apolipoprotein B48 in Transfected McA-RH7777 CellsPublished by Elsevier BV ,2003
- Galectins-3 and -7, but not Galectin-1, Play a Role in Re-epithelialization of WoundsJournal of Biological Chemistry, 2002
- Apolipoprotein Concentrations During Treatment and Recurrent Coronary Artery Disease EventsArteriosclerosis, Thrombosis, and Vascular Biology, 2000
- Site‐specific detection and structural characterization of the glycosylation of human plasma proteins lecithin:cholesterol acyltransferase and apolipoprotein D using HPLC/electrospray mass spectrometry and sequential glycosidase digestionProtein Science, 1995
- A method to increase contaminant tolerance in protein matrix‐assisted laser desorption/ionization by the fabrication of thin protein‐doped polycrystalline filmsRapid Communications in Mass Spectrometry, 1994
- Protection of low-density lipoprotein against oxidative modification by high-density lipoprotein associated paraoxonaseAtherosclerosis, 1993
- Human plasma transport of vitamin D after its endogenous synthesis.JCI Insight, 1993