CHARACTERIZATION OF A CYTOSKELETAL MATRIX ASSOCIATED WITH MYELIN FROM RAT-BRAIN
CHARACTERIZATION OF A CYTOSKELETAL MATRIX ASSOCIATED WITH MYELIN FROM RAT-BRAIN
复制标题
DOI：
10.1042/bj2600689
复制
发表时间：
1989-06-15
影响因子：
4.1
通讯作者：
BROPHY, PJ
中科院分区：
文献类型：
--
作者：
GILLESPIE, CS;WILSON, R;BROPHY, PJ
在含有Triton X-100的缓冲液中提取大鼠脑髓鞘产生了soulble馏分和不溶性残留物，富集在细胞骨架元素。洗涤剂可溶性部分和不溶性细胞骨架残留物的免疫印迹分析表明，所有的微管蛋白和超过一半的肌动蛋白被发现内的细胞骨架部分。髓鞘特异性蛋白质的分布也进行了检查，并显示，2“，3”-环核苷酸3“-磷酸水解酶（CNH 4）I和大多数髓鞘碱性蛋白质（MBP）均匀分布在两个馏分之间。相比之下，观察到大MBP（21.5 kDa）和CNE 2 II（50 kDa）几乎完全与细胞骨架部分分配。蛋白脂质蛋白主要存在于洗涤剂可溶性级分中，DM-20蛋白也是如此。蔗糖密度梯度离心的细胞骨架部分的分析表明，一个独特的子集的脂质紧密结合的细胞骨架蛋白残基。与总髓磷脂脂质相比，细胞因子相关的脂质中显著富集了神经鞘磷脂和神经鞘磷脂。这些结果表明，细胞骨架基质与多层髓鞘，并建议，这种结构可能在髓鞘发生中发挥了重要作用。
Extraction of rat brain myelin in a buffer containing Triton X-100 yielded a soulble fraction and an insoluble residue that was enriched in cytoskeletal elements. Immunoblot analysis of the detergent-soluble fraction and the insoluble cytoskeletal residue showed that all of the tubulin and more than half of the actin were found within the cytoskeletal fraction. The distribution of myelin-specific proteins was also examined, and revealed that 2'',3''-cyclic nucleotide 3''-phophohydrolase (CNPase) I and most of the myelin basic proteins (MBPs) were equally distributed between both fractions. By contrast, the large MBP (21.5 kDa) and CNPase II (50 kDa) were observed to partition almost entirely with the cytoskeletal fraction. Proteolipid protein was found predominantly in the detergent-soluble fraction, as was DM-20 protein. Analysis of the cytoskeletal fraction by sucrose-density-gradient centrifugation demonstrated that a distinct subset of lipids was tightly bound to the cytoskeletal protein residue. The cytoskeleton-associated lipid was considerably enriched in cerebroside and sphingomyelin by comparison with total myelin lipids. These results indicate that a cytoskeletal matrix is associated with multilamellar myelin, and suggest that this structure may play a fundamental role in myelinogenesis.