猪肾上腺皮质细胞膜高密度脂蛋白结合蛋白的研究
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Study of High Density Lipoprotein Binding Protein from Swine Adrenal Cortical Cell Membrane
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    为鉴定猪肾上腺皮质细胞膜是否存在高密度脂蛋白结合蛋白,并研究其特性,从新鲜猪肾上腺皮质细胞膜提取高密度脂蛋白结合蛋白,应用亲和层析法将其初步纯化。获得的可溶性蛋白经Ligandblot结合分析和Dotblot法测定,结果发现此膜蛋白与高密度脂蛋白有高度亲和力,用Westernblot法测得该蛋白的分子量为80kDa,亲和层析提取得到的结合蛋白分子量为77kDa。应用生物素-亲和素酶联免疫吸附试验测定该蛋白的结合特性:平衡解高常数(kd)=21.6μg/g,最大结合量(Bmax)=678.2mg/L蛋白。结果提示纯化的猪肾上腺皮质细胞膜高密度脂蛋白结合蛋白具有很高的亲和力,证明亲和层析法条件温和。

    Abstract:

    Aim To study the existence of high density lipoprotein (HDL) receptor in the cell membrane, and this may by provided the evidence for study of HDL receptor function.Methods HDL binding protein was prepared from fresh swine adrenal cortical cell membranes, and purified with affinity chromatography. The soluable membrane protein was analyzed by ligand blotting.Results The dot blotting showed that the membrane protein possessed high affinity with HDL, and western blotting suggested that the molecular weight (MW) of the binding protein by affinity chromatography is 77 kDa. The characters of the HDL binding protein was measured by biotin-avidin enzyme linked immunosorbent assay (BA-ELISA ): Kd= 21. 6 mg/L, Bmax= 678. 2 μg/g protein. Conclusion It is showed that the condition of affinity chromatography was moderate, and the purified swine adrenal cortical cell membrane possessed high affinity with HDL.

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黄以兰,庄庆祺,吴满平,梅美珍.猪肾上腺皮质细胞膜高密度脂蛋白结合蛋白的研究[J].中国动脉硬化杂志,1997,5(4):309~313.

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  • 收稿日期:1997-05-24
  • 最后修改日期:1997-09-12
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