Nishida H I, Nishida T
Burnsides Research Laboratory, Department of Food Science and Human Nutrition, University of Illinois, Urbana, Illinois 61801, USA.
J Biol Chem. 1997 Mar 14;272(11):6959-64. doi: 10.1074/jbc.272.11.6959.
Phospholipid transfer protein (PLTP) purified from human plasma was found to enhance the transfer of cholesterol from single bilayer vesicles containing phosphatidylcholine and cholesterol to high density lipoprotein-3. The rate of cholesterol transfer was greatly influenced by the cholesterol content of the donor vesicles. The maximal rate was observed with the vesicles containing 20-25 mol % cholesterol. This was in contrast to a progressive decline in the rate of phosphatidylcholine transfer with an increase in the cholesterol content. To determine the binding of cholesterol and phosphatidylcholine to PLTP, the mixtures of PLTP and the vesicles containing 3H-labeled phosphatidylcholine and 14C-labeled cholesterol were incubated and subjected to sucrose density gradient centrifugation. Determination of the label profiles showed that cholesterol as well as phosphatidylcholine were transferred from the vesicles to PLTP. The reversible nature of the binding was shown by the transfer of labeled cholesterol and phosphatidylcholine bound to PLTP to the acceptor vesicles or low density lipoprotein. Isothermal equilibrium binding of PLTP for cholesterol and phosphatidylcholine showed that PLTP possessed a considerably higher affinity and binding capacity for phosphatidylcholine than for cholesterol. The phosphatidylcholine binding affinity and capacity were greater when PLTP was incubated with phosphatidylcholine vesicles without cholesterol. A possible importance of PLTP-mediated cholesterol transfer in the circulation was described.
从人血浆中纯化得到的磷脂转移蛋白(PLTP)可增强胆固醇从含有磷脂酰胆碱和胆固醇的单层囊泡向高密度脂蛋白3的转移。胆固醇的转移速率受供体囊泡中胆固醇含量的显著影响。含20 - 25摩尔%胆固醇的囊泡观察到最大转移速率。这与随着胆固醇含量增加磷脂酰胆碱转移速率逐渐下降形成对比。为了确定胆固醇和磷脂酰胆碱与PLTP的结合情况,将PLTP与含有3H标记的磷脂酰胆碱和14C标记的胆固醇的囊泡混合物进行孵育,然后进行蔗糖密度梯度离心。标记物分布的测定表明胆固醇以及磷脂酰胆碱从囊泡转移到了PLTP。结合的可逆性通过将与PLTP结合的标记胆固醇和磷脂酰胆碱转移到受体囊泡或低密度脂蛋白得以体现。PLTP对胆固醇和磷脂酰胆碱的等温平衡结合表明,PLTP对磷脂酰胆碱的亲和力和结合能力比对胆固醇高得多。当PLTP与不含胆固醇的磷脂酰胆碱囊泡孵育时,磷脂酰胆碱的结合亲和力和能力更大。文中描述了PLTP介导的胆固醇转移在循环中的可能重要性。