Neurofibrillary tangle-associated collapsin response mediator protein-2 (CRMP-2) is highly phosphorylated on Thr-509, Ser-518, and Ser-522
Neurofibrillary tangle-associated collapsin response mediator protein-2 (CRMP-2) is highly phosphorylated on Thr-509, Ser-518, and Ser-522
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DOI：
10.1021/bi992323h
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发表时间：
2000-04-18
期刊：
影响因子：
2.9
通讯作者：
Ihara, Y
中科院分区：
文献类型：
--
作者：
Gu, YJ;Hamajima, N;Ihara, Y
3F3 是一种单克隆抗体，针对部分纯化的配对螺旋丝 (PHF)、强烈标记的神经原纤维缠结和一些斑块神经突，但几乎没有标记的神经纤维丝线。与对照大脑的情况相比，受阿尔茨海默氏病（AD）影响的大脑的可溶性部分中的65-kDa抗原的水平显着增加。先前通过对免疫亲和纯化的 65-kDa 抗原进行测序，将该抗原鉴定为人塌陷素反应介导蛋白 2 (hCRMP-2) [Yoshida, H.、Watanabe, A. 和 Ihara, Y. (1998) J, Biol. 1998]。化学。 273, 9761-9768]，在这里，我们证明 3F4 抗原代表 CRMP-2 的高度磷酸化形式。 3F4 反应性磷酸表位位于 hCRMP-2 的羧基末端部分，由大鼠脑提取物中的新型 45-50-kDa 蛋白激酶产生。该部分的定点诱变表明，CRMP-2 的多个位点在残基 507-522 内被差异磷酸化，并且三个位点 Thr-509、Ser-518 和 Ser-522 的磷酸化是完全 3F4 结合所必需的。 CRMP-2 碱性区域羧基端这一特殊部分的磷酸化可能在调节其活性方面发挥重要作用，并可能参与 AD 脑中退化神经突的形成。
3F3, a monoclonal antibody raised against partially purified paired helical filaments (PHFs), strongly labeled neurofibrillary tangles and some plaque neurites but barely labeled neuropil threads. The levels of the 65-kDa antigen were significantly increased in the soluble fraction of the brains affected by Alzheimer's disease (AD), as compared with that in the case of control brains. The antigen was previously identified as human collapsin response mediator protein-2 (hCRMP-2) by sequencing the immunoaffinity-purified 65-kDa antigen [Yoshida, H., Watanabe, A., and Ihara, Y. (1998) J, Biol. Chem. 273, 9761-9768], Here, we show that the 3F4 antigen represents a highly phosphorylated form of CRMP-2. The 3F4-reactive phosphoepitope was localized to the carboxyl-terminal portion of hCRMP-2, and was created by a novel 45-50-kDa protein kinase in rat brain extract. Site-directed mutagenesis of this portion showed that multiple sites of CRMP-2 are differentially phosphorylated within residues 507-522, and that phosphorylation of three sites, Thr-509, Ser-518, and Ser-522, is required for full 3F4 binding. The phosphorylation of this particular portion carboxyl-terminal to the basic region of CRMP-2 may play an important role in regulating its activity, and may be involved in the formation of degenerating neurites in AD brain.