Magee W E, Goff C W, Schoknecht J, Smith M D, Cherian K
J Cell Biol. 1974 Nov;63(2 Pt 1):492-504. doi: 10.1083/jcb.63.2.492.
Cationic liposomes composed of sphingomyelin, cholesterol, and stearylamine were prepared with horseradish peroxidase trapped inside. Stable particles were formed in which 10-12% of the enzymic activity appeared to be located at, or near, the outer surface of the liposome. Adsorption and uptake of liposomes by HeLa cells were followed cytochemically by electron microscopy and quantitated by enzyme assay and by the distribution and fate of particles labeled with [(14)C]cholesterol and [(125)I]horseradish peroxidase. The particles were adsorbed by HeLa cells at least 300 times as efficiently as was free horseradish peroxidase. Many of the particles remained at the cell surface, but numerous membrane-bound cytoplasmic inclusions were observed to contain peroxidase-staining material. In addition, many areas of the cell membrane gave a positive staining reaction. It was concluded that many particles (presumably the larger ones) did not gain access to the interior of the cells, many were phagocytized, and some enzyme was transferred to the cell membrane, perhaps as a result of fusion of the liposomal membrane with the cell membrane.
制备了由鞘磷脂、胆固醇和硬脂胺组成的阳离子脂质体,其中包裹了辣根过氧化物酶。形成了稳定的颗粒,其中10% - 12%的酶活性似乎位于脂质体的外表面或其附近。通过电子显微镜细胞化学方法追踪HeLa细胞对脂质体的吸附和摄取,并通过酶测定以及标记有[¹⁴C]胆固醇和[¹²⁵I]辣根过氧化物酶的颗粒的分布和命运进行定量。这些颗粒被HeLa细胞吸附的效率至少是游离辣根过氧化物酶的300倍。许多颗粒留在细胞表面,但观察到大量膜结合的细胞质内含物含有过氧化物酶染色物质。此外,细胞膜的许多区域呈现阳性染色反应。得出的结论是,许多颗粒(可能是较大的颗粒)无法进入细胞内部,许多被吞噬,并且一些酶被转移到细胞膜,这可能是脂质体膜与细胞膜融合的结果。