Characterization of Interactions between the Neurofilament Triplet Proteins by the Yeast Two-hybrid System*
Characterization of Interactions between the Neurofilament Triplet Proteins by the Yeast Two-hybrid System*
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酵母双杂交系统表征神经丝三联体蛋白之间的相互作用*
DOI：
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发表时间：
1996
影响因子：
4.8
通讯作者：
R. Liem
中科院分区：
文献类型：
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作者：
C. Leung;R. Liem
在成年轴突中，神经丝（NFs）是由低（NFL）、中（NFM）和高（NFH）分子量的神经丝三重体蛋白（NFTPs）形成的异聚合物。这三种蛋白都具有基本的中间丝蛋白三方结构，包括一个短的氨基端头区、一个约310个氨基酸的α-螺旋棒区和一个可变长度的羧基端尾区。体外聚合研究表明，只有NFL可以组装成10nm的均聚长丝。包括NFs在内的中间细丝的组装始于涉及两个分子的α-螺旋棒结构域的盘绕二聚体的形成。为了确定NFTPs的同型二聚体还是异源二聚体是NFs组装的首选中间体，我们使用酵母双杂交系统研究了不同NFTPs之间的相互作用。通过监测lacZ报告基因产物的活性，我们能够证明NFL与NFL、NFM或NFH的相互作用强于NFM与NFM或NFH的相互作用以及NFH与NFH的相互作用。这些结果表明NFM和NFH比同型二聚体更容易与NFL形成异源二聚体，这与NFM和NFH在体外和体内都不能自聚合是一致的。
In the adult axon, the neurofilaments (NFs) are heteropolymers formed from the low (NFL), middle (NFM), and high (NFH) molecular weight neurofilament triplet proteins (NFTPs). All three proteins have the basic intermediate filament protein tripartite structure, which consists of a short amino-terminal head region, an α-helical rod region of ∼310 amino acids, and a carboxyl-terminal tail region of variable length. In vitro polymerization studies have shown that only NFL can assemble into homopolymeric 10-nm filaments. The assembly of intermediate filaments, including the NFs, begins with the formation of a coiled-coil dimer involving the α-helical rod domains of two molecules. In order to determine whether homodimers or heterodimers of NFTPs are the preferred intermediates in the assembly of NFs, we have used the yeast two-hybrid system to study the interactions between the different NFTPs. By monitoring the activity of the lacZ reporter gene product, we are able to show that the interactions of NFL with NFL, NFM, or NFH are stronger than the interactions of NFM with NFM or NFH and the interaction of NFH with NFH. These results imply that NFM and NFH are more likely to form heterodimers with NFL than homodimers and are consistent with the inability of NFM and NFH to self-polymerize in vitro and in vivo.
截短的神经丝蛋白对转染的成纤维细胞内源性中间丝的影响。
影响因子：
4
作者：
Chin,SS;Macioce,P;Liem,RK
通讯作者：
Liem,RK
牛神经丝的抗体标记：对神经丝侧臂结构的影响。
影响因子：
3
作者：
Mulligan,L;Balin,BJ;Lee,VM;Ip,W
通讯作者：
Ip,W
大鼠神经丝蛋白 NF-L 和 NF-M 在转染的非神经元细胞中的表达。
影响因子：
6.6
作者：
Chin,SS;Liem,RK
通讯作者：
Liem,RK