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首页> 外文期刊>Journal of Lipid Research >Evidence for the presence of lipid-free monomolecular apolipoprotein A-1 in plasma
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Evidence for the presence of lipid-free monomolecular apolipoprotein A-1 in plasma

机译：血浆中存在无脂单分子载脂蛋白A-1的证据

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The first step in reverse cholesterol transport is a process by which lipid-free or lipid-poor apoA-1 removes cholesterol from cells through the action of ATP binding cassette transporter A1 at the plasma membrane. However the structure and composition of lipid-free or -poor apoA-1 in plasma remains obscure. We previously obtained a monoclonal antibody (MAb) that specifically recognizes apoA-1 in pre{beta}1-HDL, the smallest apoA-1-containing particle in plasma, which we used to establish a pre{beta}1-HDL ELISA. Here, we purified pre{beta}1-HDL from fresh normal plasma using said antibody, and analyzed the composition and structure. ApoA-1 was detected, but neither phospholipid nor cholesterol were detected in the purified pre{beta}1-HDL. Only globular, not discoidal, particles were observed by electron microscopy. In nondenaturing PAGE, no difference in the mobility was observed between the purified pre{beta}1-HDL and original plasma pre{beta}1-HDL, or between the pre{beta}1-HDL and lipid-free apoA-1 prepared by delipidating HDL. In sandwich ELISA using two anti-pre{beta}1-HDL MAbs, reactivity with intact plasma pre{beta}1-HDL was observed in ELISA using two MAbs with distinct epitopes but no reactivity was observed in ELISA using a single MAb, and the same phenomenon was observed with monomolecular lipid-free apoA-1. These results suggest that plasma pre{beta}1-HDL is lipid-free monomolecular apoA-1.
机译：胆固醇逆向转运的第一步是一个过程，通过该过程，无脂或贫脂的apoA-1通过质膜上的ATP结合盒转运蛋白A1的作用从细胞中去除胆固醇。然而，血浆中无脂或贫apoA-1的结构和组成仍然不清楚。我们先前获得了一种单克隆抗体（MAb），该抗体可特异性识别血浆中最小的载脂蛋白A-1的前β1-HDL中的apoA-1，我们用来建立前β1-HDLELISA。在此，我们使用所述抗体从新鲜的正常血浆中纯化了前β1-HDL，并分析了其组成和结构。检测到ApoA-1，但在纯化的preβ1-HDL中未检测到磷脂和胆固醇。通过电子显微镜仅观察到球形而非盘状颗粒。在非变性PAGE中，在纯化的preβ1-HDL和原始血浆preβ1-HDL之间，或在preβ1-HDL和制备的无脂质apoA-1之间，没有观察到迁移率的差异。通过使HDL脱脂。在使用两个抗preβ1-HDLMAb的三明治ELISA中，在使用两个具有不同表位的单克隆抗体的ELISA中观察到与完整血浆preβ1-HDL的反应性，但是在使用单个MAb的ELISA中未观察到反应性。用单分子无脂质apoA-1观察到相同的现象。这些结果表明血浆前β1-HDL是不含脂质的单分子apoA-1。

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