PURIFICATION AND CHARACTERIZATION OF A BOVINE ACETYL LOW-DENSITY LIPOPROTEIN RECEPTOR
PURIFICATION AND CHARACTERIZATION OF A BOVINE ACETYL LOW-DENSITY LIPOPROTEIN RECEPTOR
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DOI：
10.1073/pnas.85.23.9238
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发表时间：
1988-12-01
影响因子：
11.1
通讯作者：
KRIEGER, M
中科院分区：
文献类型：
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作者：
KODAMA, T;REDDY, P;KRIEGER, M
乙酰基低密度脂蛋白（LDL）受体在巨噬细胞和某些内皮细胞上表达，并介导培养物中巨噬细胞-泡沫细胞的形成。从牛肝膜中部分纯化了220-kDa的乙酰基低密度脂蛋白结合蛋白，并用于制备特异性单克隆抗体。220-kDa的蛋白免疫沉淀由该抗体保留结合活性，和抗体被用来检测这种蛋白质在细胞内衬牛肝窦和培养的牛肺泡巨噬细胞的表面上。在人单核细胞系THP-1中，乙酰LDL受体活性和220-kDa乙酰LDL结合蛋白的表达在分化成巨噬细胞样状态后由佛波酯诱导平行地显著诱导。配体的特异性，组织和细胞类型的特异性，和共诱导的数据表明，这220 kDa的细胞表面结合蛋白可能是一个受体，介导乙酰LDL内吞作用。使用单克隆抗体亲和层析从牛肺膜纯化238，000倍至接近同质的220-kDa蛋白质是含有天冬酰胺连接的碳水化合物链的77-kDa亚基的三聚体。
The acetyl low density lipoprotein (LDL) receptor is expressed on macrophages and some endothelial cells and mediates macrophage-foam cell formation in culture. A 220-kDa acetyl LDL binding protein was partially purified from bovine liver membranes and was used to make a specific monoclonal antibody. The 220-kDa protein immunoprecipitated by this antibody retained binding activity, and the antibody was used to detect this protein in cells lining bovine liver sinusoids and on the surface of cultured bovine alveolar macrophages. In the human monocytic cell line THP-1, the expression of both acetyl LDL receptor activity and a 220-kDa acetyl LDL binding protein were dramatically induced in parallel after differentiation to a macropahge-like state induced by phorbol ester. The ligand specificity, tissue and cell-type specificity, and coinduction data indicated that this 220-kDa cell-surface binding protein is probably a receptor that mediates acetyl LDL endocytosis. The 220-kDa protein, which was purified 238,000-fold from bovine lung membranes to near homogeneity using monoclonal antibody affinity chromatography, is a trimer of 77-kDa subunits that contain asparagine-linked carbohydrate chains.