Determination of Lipid Transfer Inhibitor Protein Activity in Human Lipoprotein-Deficient Plasma

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Lipid transfer protein (LTP) activity is modulated by a distinct plasma protein termed lipid transfer inhibitor protein (LTIP). The objective of this study was to establish an assay for LTIP that could be used to quantify its activity in lipoprotein-deficient plasma. A straightforward heating protocol (56°C for 60 minutes) was found to inactivate more than 90% of LTIP activity. The responses of individual lipoprotein-deficient plasma samples to this heating procedure were unique. Among normolipidemic donors, inactivation of LTIP caused a 230% to 600% increase in LTP activity. Essentially all measurable transfer activity in native and heated samples was inhibited by an antibody to LTP. Whole-plasma samples from these donors were spiked with radiolabeled lipoproteins to measure the rates of lipid transfer among the major lipoprotein classes. In general, plasma lipid transfer rates were negatively correlated with LTIP activity in these samples. However, the decrease in lipid transfers from very-lowdensity lipoprotein (VLDL) to low-density lipoprotein (LDL) and from LDL to VLDL was from 2.4- to 5.1-fold greater than in the transfers from VLDL to high-density lipoprotein (HDL) or from HDL to VLDL. In these samples, the molecular weight of HDL2 was negatively correlated with LTIP activity. Thus, LTIP activities among normolipidemic individuals were observed to vary severalfold; compared with other lipoprotein transfers, higher LTIP activities were associated with a relative reduction in LDL-VLDL lipid transfer events.

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