Phosphorylatin of NPC1 by Cdk5 in Niemann-Pick diseases.

尼曼-皮克病中 Cdk5 对 NPC1 的磷酸化。

• 批准号: 18500243
• 负责人: TOMIZAWA Kazuhito
• 金额: $ 2.57万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18500243/
• 关键词: lipid transport; membrane transport; Cdk5; cyclin; Niemann-Pick diseases; neurodegeneration; NPC1; サイクリン依存性リン酸化酵素

Niemann-Pick type Cl (NPC1) disease is an autosomal-recessive cholesterol-storage disorder characterized by liver dysfunction, hepatosplenomegaly, and progressive neurodegeneration. At the cellular level, NPC1 disease is characterized by an accumulation of cholesterol within late endosomes/lysosomes derived from both endogenously synthesized cholesterol and endocytosis of lipoprotein-derived cholesterol through the coated-pit pathway. Mutations of NPC1 gene are found about 95 % in the patients with NPC1 disease. The mutations are observed both intracellular domain and the binding domain with lipid.The aim of present study is to examine the effect of phosphorylation of the intracellular domain of NPC1 on abnormal intracellular cholesterol homeostasis in Niemann-Pick diseases. Mutant NPC1 seen in the patients with Niemann-Pick diseases was phosphorylated by cyclin-dependent Iciness 5 (Cdk5) in vitro. We identified the phosphorylation sites. Cdk5 phosphorylated NPC1 at Ser320. Phospho-specific antibody, which recognizes the Cdk5-dependent phosphorylation of NPC1 reveals that mutant NPC1 in the fibroblasts in patients with Niemann-Pick diseases is phosphorylated by Cdk5. Moreover, the phosphorylation was inhibited by Cdk5 inhibitors. In contrast, the phosphorylation was not detected in wild-type NPC1 expressing fibroblasts. Finally, the phosphorylation inhibited the change of NPC1 localization to mitochondria.

Niemann-Pick型Cl （NPC1）病是一种常染色体隐性胆固醇储存疾病，以肝功能障碍、肝脾肿大和进行性神经变性为特征。在细胞水平上，NPC1疾病的特征是胆固醇在内源性合成胆固醇和通过包被窝途径内吞脂蛋白源性胆固醇的晚期内体/溶酶体中积累。NPC1基因突变在NPC1病患者中约占95%。细胞内结构域和脂质结合结构域均观察到突变。本研究旨在探讨NPC1细胞内结构域磷酸化对尼曼-匹克病细胞内异常胆固醇稳态的影响。在Niemann-Pick病患者中发现的突变体NPC1在体外被细胞周期蛋白依赖性Iciness 5 （Cdk5）磷酸化。我们确定了磷酸化位点。Cdk5磷酸化NPC1的Ser320位点。磷酸化特异性抗体识别Cdk5依赖的NPC1磷酸化，表明Niemann-Pick病患者成纤维细胞中的突变体NPC1被Cdk5磷酸化。此外，磷酸化被Cdk5抑制剂抑制。相比之下，在表达NPC1的野生型成纤维细胞中未检测到磷酸化。最后，磷酸化抑制了NPC1在线粒体定位的变化。

项目成果

A Cdk5 inhibitor enhances induction of insulin secretion by exendin-4 both in vitro and in vivo.

Cdk5 抑制剂可在体外和体内增强 Exendin-4 对胰岛素分泌的诱导。

• 发表时间: 2007
• 期刊: J. Physiol. Sci. 57
• 作者: Kitani;K.
• 通讯作者: K.

Novel protein transduction method by using 11R. An effectiv new drug delivery system fro the treatment of cerebrovascular diseases.

使用11R的新型蛋白质转导方法。

• 发表时间: 2007
• 期刊: Stroke (印刷中)
• 作者: Ogawa;N. et al.
• 通讯作者: N. et al.

Major Cdk5-dependent phosphorylation sites of amphiphysin 1are implicated in the regulation of the membrane binding and endocytosis.

两性蛋白 1 的主要 Cdk5 依赖性磷酸化位点参与膜结合和内吞作用的调节。

• 发表时间: 2007
• 期刊: Journal of Neurochemistry (印刷中)
• 作者: Liang;S. et al.
• 通讯作者: S. et al.

Truncations of amphiphysin I by calpain inhibit vesicle endocytosis during neural hyperexcitation

• DOI: 10.1038/sj.emboj.7601741
• 发表时间: 2007-06-20
• 期刊: EMBO JOURNAL
• 影响因子: 11.4
• 作者: Wu, Yumei;Liang, Shuang;Tomizawa, Kazuhito
• 通讯作者: Tomizawa, Kazuhito

Mice overexpressing dominant negative Cdk5 in the pancreatic beta-cell show the phenotype of type 1 diabetes mellitus

胰腺β细胞中过度表达显性失活Cdk5的小鼠表现出1型糖尿病的表型

• 发表时间: 2008
• 作者: Yoshihiro;Ohtani;et. al.
• 通讯作者: et. al.