Structure and composition of the cytoskeleton of nucleated erythrocytes I. The presence of microtubule-associated protein 2 in the marginal band
Structure and composition of the cytoskeleton of nucleated erythrocytes I. The presence of microtubule-associated protein 2 in the marginal band
复制标题
有核红细胞细胞骨架的结构和组成 I. 边缘带微管相关蛋白 2 的存在
DOI：
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发表时间：
1980
影响因子：
7.8
通讯作者：
Kay Dickersin
中科院分区：
文献类型：
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作者：
R. Sloboda;Kay Dickersin
有核红细胞的边缘带（MB）由一束微管组成，环绕质膜下的细胞。当细胞在含有Triton X-100的缓冲液中裂解时，MB微管保持完整，细胞核通过称为trans-MB材料的丝状网络悬浮在细胞中心，该丝状网络将细胞核连接到外周MB。当使用鸡脑微管相关蛋白2（MAP 2）的抗体制备这些裂解细胞用于间接免疫荧光时，MB产生强烈染色;在细胞核或trans-MB物质占据的区域中无明显染色。对照证明染色具有特异性，因为单独使用荧光山羊抗兔血清或使用非免疫血清作为第一抗体时未发生染色。此外，MB的荧光不受用纯化的微管蛋白预处理免疫血清的影响，但通过用纯化的MAP 2预处理免疫血清防止染色。为了确定哪种MB蛋白组分导致阳性免疫荧光结果，在分离的细胞骨架的蛋白组分通过SDS-聚丙烯酰胺凝胶解析后，使用125 I-蛋白A染色。用SDS凝胶电泳法检测抗血清与纯化的鸡脑微管MAP 2的反应。细胞骨架蛋白的结果表明，抗血清仅与高分子量蛋白反应，所述高分子量蛋白的分子量与鸡脑MAP 2的分子量相似，但不相同。因此，可以得出结论，具有类似于鸡脑MAP 2的抗原特性的蛋白质是MB的组分。结果进行了讨论的MAP 2在MB中的可能功能。
The marginal band (MB) of nucleated erythrocytes is composed of a bundle of microtubules that encircles the cell immediately beneath the plasma membrane. When cells are lysed in buffer containing Triton X-100 the MB microtubules remain intact, and the nucleus remains suspended at the cell center by a filamentous network called the trans-MB material that connects the nucleus to the peripheral MB. When these lysed cells are prepared for indirect immunofluorescence by use of an antibody to chick brain microtubule-associated protein 2 (MAP 2), intense staining of the MB results; no staining is evident in the areas occupied by the nucleus or the trans-MB material. Controls demonstrate that the staining is specific, because no staining occurs with fluorescent goat antirabbit serum alone or when nonimmune serum is used as the first antibody. Furthermore, the fluorescence of the MB is not affected by pretreatment of the immune serum with purified tubulin, but staining is prevented by pretreatment of the immune serum with purified MAP 2. To determine which protein component of the MB was responsible for the positive immunofluorescence results, 125I-protein A staining was used after the protein components of the isolated cytoskeleton had been resolved by SDS-polyacrylamide gels. Controls showed that the antiserum could react on SDS gels with MAP 2 from purified chick brain microtubules. The results with the cytoskeletal proteins demonstrated that the antiserum reacted only with a high molecular weight protein having a molecular weight similar, but not identical, to that of chick brain MAP 2. Thus, it is concluded that a protein with antigenic characteristics similar to those of chick brain MAP 2 is a component of the MB. The results are discussed in terms of the possible function of MAP 2 in the MB.