Spectrin rearrangement early in erythrocyte ghost endocytosis.
Spectrin rearrangement early in erythrocyte ghost endocytosis.
复制标题
DOI：
10.1083/jcb.82.3.654
复制
发表时间：
1979-09
期刊：
影响因子：
--
通讯作者：
Schrier SL
中科院分区：
文献类型：
--
作者：
Hardy B;Bensch KG;Schrier SL
在白鬼诱导的内吞空泡中发现了幽灵蛋白的耗尽，因此提出它们是由红细胞膜中无幽灵蛋白的区域产生的。为了跟踪胞吞过程中spectrin分布的变化，制备了亲和纯化的兔抗spectrin抗体。定量技术被开发用于使用高度特异性的125I-F(ab')2抗光谱蛋白，这些表明，在液泡出现之前，通过相显微镜评估，免疫反应性光谱蛋白有可重复的减少。为了确定这种谱蛋白减少是局部或漫射谱蛋白丢失还是谱蛋白重排，使用透射电子显微镜对兔抗谱蛋白和铁蛋白偶联山羊抗兔免疫球蛋白处理的样品进行了形态学研究。这些研究表明，内吞作用发生之前，产生了广泛的无谱区，由离散的含谱区隔开。用碱性磷酸酶对鬼影进行预处理，可阻断各种形式的内吞作用，防止无鬼影区产生。因此，我们提出在内吞诱导剂的推动下，磷酸化的谱蛋白被重新分配，从而产生无谱区，在无谱区形成内吞液泡并融合。
The endocytic vacuoles induced in white ghosts were found to be depleted of spectrin and therefore it was proposed that they arose from spectrin-free areas in the erythrocyte membrane. To follow changes in spectrin distribution during endocytosis, affinity-purified rabbit antispectrin antibodies were produced. Quantitative techniques were developed for the use of a highly specific 125I-F(ab')2 antispectrin, and these showed that before the appearance of vacuoles, as assessed by phase microscopy, there was a reproducible decrease in immunoreactive spectrin. To determine whether this spectrin decrease represented a local or diffuse spectrin loss or a spectrin rearrangement, morphologic studies were undertaken using transmission electron microscopy on samples treated with rabbit antispectrin and ferritin-conjugated goat anti-rabbit immunoglobulin. These studies showed that endocytosis was preceded by the creation of extensive spectrin-free areas separated by discrete spectrin-containing zones. Pretreatment of ghosts with alkaline phosphatase blocked all forms of endocytosis and prevented the creation of spectrin-free areas. Therefore, it is proposed that under the impetus of endocytosis inducers, phosphorylated spectrin is redistributed so that spectrin-free zones are created, and that endocytic vacuoles form and fuse in spectrin-free areas.