Membrane rafts may act as platforms for membrane protein signalling. Rafts have also been implicated in the sorting of membrane components during membrane budding. We have studied by fluorescence microscopy cross-linking of ganglioside GM1 in the human erythrocyte membrane, and how membrane proteins CD47 and CD59 distribute in GM1 patched discoid cells and calcium-induced echinocytic cells. Patching of ganglioside(M1) (GM1) by cholera toxin subunit B (CTB) plus anti-CTB resulted in the formation of usually 40-60 GM1 patches distributed over the membrane in discoid erythrocytes. Pre-treatment of erythrocytes with methyl-beta-cyclodextrin abolished GM1 patching. GM1 patching was insensitive to pre-fixation (paraformaldehyde) of cells. Patching of GM1 did not affect the discoid shape of erythrocytes. Membrane proteins CD47 and CD59 did not accumulate into GM1 patches. No capping of patches occurred. GM1 accumulated in calcium-induced echinocytic spiculae. Also CD59, but not CD47, accumulated in spiculae. However, CD59 showed a low degree of co-localization with GM1 and frequently accumulated in different spiculae than GM1. In conclusion, our study describes a novel method for examining properties and composition of rafts. The study characterizes raft patching in the human erythrocyte membrane and emphasizes the mobility and 'echinophilicity' of GM1. Glycosyl phosphatidylinositol-anchored CD59 was identified as a mobile 'echinophilic' but 'raftophobic(GM1)' protein. Largely immobile CD47 showed no segregation.

中文翻译：

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人类红细胞中神经节苷脂（M1）的修补-修补和弯曲的膜中CD47和CD59的分布。

膜筏可以充当膜蛋白信号传导的平台。在膜出芽过程中，筏也与膜成分的分类有关。我们已通过荧光显微镜研究了神经节苷脂GM1在人红细胞膜中的交联，以及膜蛋白CD47和CD59如何在GM1修补的盘状细胞和钙诱导的嗜酸性细胞中分布。霍乱毒素亚基B（CTB）加抗CTB修补神经节苷脂（M1）（GM1）导致形成通常40-60 GM1斑块分布在盘状红细胞的膜上。用甲基-β-环糊精进行的红细胞预处理消除了GM1补丁。GM1修补对细胞的预固定（低聚甲醛）不敏感。修补GM1不会影响红细胞的盘状形状。膜蛋白CD47和CD59没有积累到GM1斑块中。没有补丁的上限。GM1积累在钙诱导的棘突性针状刺中。针状体中也积累了CD59，但没有CD47。但是，CD59与GM1的共定位程度较低，并且经常在与GM1不同的针状体中积聚。总之，我们的研究描述了一种新颖的方法来检查木筏的性质和成分。这项研究的特点是筏在人类红细胞膜上的修补，并强调了GM1的迁移性和“嗜酸性”。糖基磷脂酰肌醇锚定的CD59被确定为一种可移动的“嗜酸性”但“ raftophobic（GM1）”蛋白。高度固定的CD47没有显示偏析。针状体中也积累了CD59，但没有CD47。但是，CD59与GM1的共定位程度较低，并且经常在与GM1不同的针状体中积聚。总之，我们的研究描述了一种新颖的方法来检查木筏的性质和成分。这项研究的特点是筏在人类红细胞膜上的修补，并强调了GM1的迁移性和“嗜酸性”。糖基磷脂酰肌醇锚定的CD59被确定为一种可移动的“嗜酸性”但“ raftophobic（GM1）”蛋白。高度固定的CD47没有显示偏析。针状体中也积累了CD59，但没有CD47。但是，CD59与GM1的共定位程度较低，并且经常在与GM1不同的针状体中积聚。总之，我们的研究描述了一种新颖的方法来检查木筏的性质和成分。这项研究的特点是筏在人类红细胞膜上的修补，并强调了GM1的迁移性和“嗜酸性”。糖基磷脂酰肌醇锚定的CD59被鉴定为可移动的“嗜酸性”但“反亲（GM1）”蛋白。高度固定的CD47没有显示偏析。这项研究的特点是筏在人类红细胞膜上的修补，并强调了GM1的迁移性和“嗜酸性”。糖基磷脂酰肌醇锚定的CD59被确定为一种可移动的“嗜酸性”但“ raftophobic（GM1）”蛋白。高度固定的CD47没有显示偏析。这项研究的特点是筏在人类红细胞膜上的修补，并强调了GM1的迁移性和“嗜酸性”。糖基磷脂酰肌醇锚定的CD59被确定为一种可移动的“嗜酸性”但“ raftophobic（GM1）”蛋白。高度固定的CD47没有显示偏析。