Neurofilament proteins of rat peripheral nerve and spinal cord.
Neurofilament proteins of rat peripheral nerve and spinal cord.
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DOI：
10.1083/jcb.78.3.653
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发表时间：
1978-09
影响因子：
7.8
通讯作者：
Freeman, L A
中科院分区：
文献类型：
--
作者：
Schlaepfer, W W;Freeman, L A
通过渗透压休克将完整的神经丝平行地从大鼠周围神经和脊髓分离到含有二价阳离子螯合剂的低渗介质中。分离的神经丝经过 0.1 M NaCl 多次离心洗涤和分离。通过负染色技术在洗涤的沉淀中鉴定出丰富的完整神经丝。它们起源于神经丝，并通过免疫电子显微镜证实。用 8 M 尿素从脂质和膜成分中提取洗涤后的神经丝。对十二烷基硫酸钠凝胶上的神经丝分离物的分析表明，200,000、150,000 和 69,000 mol wt 的蛋白质是源自大鼠外周和中枢神经系统的完整神经丝的主要成分。这些相同的蛋白质在两种来源的整个组织匀浆中都被鉴定出来，并且在完整神经丝的分离过程中变得丰富。分离过程中产生了 64,000 mol wt 的少量组分。其他蛋白质被鉴定为污染物。少量带有微管蛋白和肌动蛋白电泳迁移的蛋白质保留在神经丝分离物中。
Intact neurofilaments were isolated in parallel from rat peripheral nerve and spinal cord by osmotic shock into hypotonic media containing divalent cation chelators. Isolated neurofilaments were washed and separated by multiple centrifugations in 0.1 M NaCl. Abundant intact neurofilaments were identified in the washed pellets by negative staining techniques. Their origin from neurofilaments was confirmed by immune electron microscopy. Washed neurofilaments were extracted from lipid and membranous components with 8 M urea. Analyses of neurofilament isolates on sodium dodecyl sulfate gels showed that proteins of 200,000, 150,000, and 69,000 mol wt were the major components of intact neurofilaments derived from rat peripheral and central nervous systems. These same proteins were identified in whole tissue homogenates of both sources and became enriched during the isolation of intact neurofilaments. A minor component of 64,000 mol wt arose during isolation. Other proteins were identified as contaminants. Small amounts of proteins with electrophoretic migration of tubulin and actin remain in neurofilament isolates.