IDENTIFICATION OF CA2+-ACTIVATED NEUTRAL PROTEASE IN THE PERIPHERAL-NERVE AND ITS EFFECTS ON NEUROFILAMENT DEGENERATION
IDENTIFICATION OF CA2+-ACTIVATED NEUTRAL PROTEASE IN THE PERIPHERAL-NERVE AND ITS EFFECTS ON NEUROFILAMENT DEGENERATION
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DOI：
10.1111/j.1471-4159.1983.tb08072.x
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发表时间：
1983-01-01
影响因子：
4.7
通讯作者：
TOYOKURA, Y
中科院分区：
文献类型：
--
作者：
KAMAKURA, K;ISHIURA, S;TOYOKURA, Y
将大鼠坐骨神经在5种不同的培养液中孵育。神经丝三联蛋白(200K、160K和68K mW)在含钙离子的介质中消失，并被E--c[N-(N-[L-3-carboxyoxirane-2-carbonyl]-L-leucyl)-isoamylamine]或亮肽素抑制。在周围神经中存在一种酶，其性质类似于钙激活的中性蛋白酶(CANP)。从大鼠坐骨神经提取CANP粗品。钙调素酶活力完全恢复(0.129.+-。在周围神经可溶部分(粗CANP)中加入0.008-50%饱和(NH_4)_2SO_4盐析出的沉淀物中。以核因子为底物对粗CANP的性质进行了检测，发现其性质与从骨骼肌中提取的CANP相似。用免疫复制法对从大鼠骨骼肌中提取的CANP进行粗品鉴定。在两个CANP中均检测到与73K对应的条带。
Rat sciatic nerve segments were incubated in 5 different media. Disappearance of neurofilament (NF) triplet proteins (200K [kilodalton], 160K and 68K MW) occurred in medium containing Ca2+ and was inhibited by the addition of E-64-c [N-(N-[L-3-carboxyoxirane-2-carbonyl]-L-leucyl)-isoamylamine] or leupeptin. The presence in the peripheral nerve of an enzyme whose properties are similar to those of Ca2+-activated neutral protease (CANP) is suggested. The extraction of crude CANP from rat sciatic nerve was performed. CANP activity was completely recovered (0.129 .+-. 0.008 U/g) in the precipitate salted out by the addition of 0-50% saturated (NH4)2SO4 to the soluble fraction of the peripheral nerve (crude CANP). Properties of the crude CANP were examined using NF as a substrate and were found to be similar to those of the CANP extracted from skeletal muscle. Identification of the crude CANP with the CANP extracted from rat skeletal muscle was performed using the immunoreplica method. Bands corresponding to 73K were detected in both CANP.