Bottum K, Jonas A
Department of Biochemistry, College of Medicine, University of Illinois at Urbana-Champaign 61801, USA.
Biochemistry. 1995 May 30;34(21):7264-70. doi: 10.1021/bi00021a043.
Cholesterol spontaneously transfers from low density lipoproteins (LDL) to high density lipoproteins (HDL). This transfer is important physiologically as it supplies the major portion of cholesterol for the lecithin:cholesterol acyltransferase reaction and is one mechanism for the reduction of atherogenic LDL cholesterol. The objective of this work was to examine the properties of both HDL and LDL which modulate cholesterol transfer, as well as to obtain the relevant kinetic constants for the transfer at concentrations of lipoproteins approaching those existing in vivo. To examine the effects of HDL structural parameters on cholesterol transfer, we prepared reconstituted HDL particles with saturated or unsaturated phospholipid, with apolipoprotein AI or apolipoprotein AII, with increasing size and phospholipid content, and with increasing initial contents of cholesterol. We also prepared five LDL subfractions of variable density and size. The kinetics of cholesterol mass transfer were measured by incubating LDL with rHDL at 37 degrees C, separating the lipoproteins by dextran sulfate/Mg2+ precipitation of LDL at timed intervals, and analyzing rHDL cholesterol content. The cholesterol content of rHDL at equilibrium, Ceq, and the half-time for transfer, t1/2, as well as the ratio of the lipid surface areas of LDL to rHDL were used in the analysis of the kinetic data by the aqueous diffusion model for lipid transfer developed by Nichols and Pagano [(1982) Biochemistry 21, 1720-1726]. The only variables that significantly affect the C(eq) and/or t1/2 are the phospholipid content and composition of the rHDL and the size or density of the LDL particles.(ABSTRACT TRUNCATED AT 250 WORDS)
胆固醇会自发地从低密度脂蛋白(LDL)转移至高密度脂蛋白(HDL)。这种转移在生理上很重要,因为它为卵磷脂胆固醇酰基转移酶反应提供了大部分胆固醇,并且是降低致动脉粥样硬化的LDL胆固醇的一种机制。这项工作的目的是研究调节胆固醇转移的HDL和LDL的特性,以及在脂蛋白浓度接近体内存在浓度时获得转移的相关动力学常数。为了研究HDL结构参数对胆固醇转移的影响,我们制备了具有饱和或不饱和磷脂、载脂蛋白AI或载脂蛋白AII、尺寸和磷脂含量增加以及初始胆固醇含量增加的重组HDL颗粒。我们还制备了五个密度和尺寸可变的LDL亚组分。通过在37℃下将LDL与重组HDL一起孵育、在不同时间间隔通过硫酸葡聚糖/Mg2+沉淀LDL来分离脂蛋白并分析重组HDL胆固醇含量,来测量胆固醇质量转移的动力学。通过Nichols和Pagano [(1982年)《生物化学》21, 1720 - 1726] 开发的脂质转移水相扩散模型,将重组HDL在平衡时的胆固醇含量Ceq、转移的半衰期t1/2以及LDL与重组HDL的脂质表面积比用于动力学数据分析。唯一显著影响C(eq)和/或t1/2的变量是重组HDL的磷脂含量和组成以及LDL颗粒的尺寸或密度。(摘要截取自250字)