Identification and quantification of proteins participating in apoA-1 and apoB-100 mediated cholesterol transport by field-flow fractionation and LC-MS/MS analysis
Zsuzsanna Kuklenyik
Centers for Disease Control and Prevention, USA
: J Diagnos Tech Biomed Anal
Abstract
Lipid homeostasis in vivo is mediated by several lipoprotein sub-species, commonly classified as high density and low density lipoproteins (HDL and LDL). The two common apolipoproteins, ApoA-1 and ApoB-100 are associated with numerous exchangeable lipids and proteins in various compositions. Because of their number and complexity, the analysis of lipoprotein subspecies in plasma by bottom-up proteomics approaches requires pre-analytical physical fractionation. Asymmetric flow-field flow fraction (AF4) is a gentle separation technique based on hydrodynamic size. AF4 allows size separation of intact lipoprotein subspecies and fraction collection. LC-MS/MS analysis of the individual fractions can provide unique concentration versus hydrodynamic size profiles for each protein constituent. Deconvolution of the concentration versus size profiles of ApoA-1 and ApoB-100 allows quantitative deduction of the particle number of lipoprotein sub-species. The probability of associations of proteins with ApoA-1 and ApoB-100 containing sub-species was evaluated based on size profile overlap with those of Apo-A-1 or ApoB-100 sub-species, size derived maximum possible total molecular weight, and protein-ApoA-1 or protein-ApoB-100 concentration correlations measured in multiple serum samples with wide range of cholesterol and triglyceride levels. Based on these criteria proteins can be included or excluded as participants of ApoA-1 and ApoB-100 mediated lipid metabolism pathways.
Biography
Zsuzsanna Kuklenyik has completed her master degree in Chemical Engineering at Technical University of Budapest, and her PhD degree at Emory University of Atlanta Georgia, where she also conducted Postdoctoral studies. Currently, she is a Senior Research Scientist in the Biological Mass Spectrometry Laboratory at the Centers for Disease Control and Prevention in Atlanta. She has published more than 40 papers in reputed journals on wide range of applications of hyphenated chromatographic techniques and mass spectrometry, such as biomonitoring of environmental chemicals, analysis of pre-exposure prophylactic drugs against HIV, biological toxins, and more recently, lipoproteins.
