Stabilization of ion channel protein by regulation of proteasome and its application to the individualizing atrial fibrillation.

通过蛋白酶体调节稳定离子通道蛋白及其在个体化心房颤动中的应用。

• 批准号: 15590747
• 负责人: HISATOME Ichiro
• 金额: $ 2.24万
• 依托单位: National University Corporation Tottori University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590747/
• 关键词: atrial fibrillation; ion channel; ubiquitin-proteasome; Na^+ channel blockers; SNPs; ユビキチン・プロテアソーム; Na channel 阻害剤; 蛋白安定化; Kv1.5; HSP70; ユビキチン化; 細胞内トラフィック; 小胞体品質管理機構

Background : Previously we reported that ion channels responsible for chronic atrial fibrillation have been degraded by ubiquitin-proteasome, which was modification by antiarrhythmic agents. In the current study, we elucidated its underlying mechanism and application to the individualizing therapy for atrial fibrillation.Methods and Results : Na^+ channel blockers, but not either Ca^<2+> antagonists or K^+ channel blockers, blocked 20S proteasome activity to stabilize the short-lived proteins including p53,IKK2 and Kv1.5 assessed by pulse-chase analysis, immnofluorescence and patch clamp techniques in transfected COS cells. To confirm that p53 stabilization by Na^+ channel blockers was secondary to proteasome inhibition, we examined whether Mdm2 (C438A) could block the drug effects. When co-expressed with Mdm2 (C438A), p53-FLAG was not at all ubiquitinated even when cells were treated with MG132 and this drug failed to increase the p53-FLAG protein level. Likewise, p53-FLAG was not ubiquitinated in cells treated with Na^+ channel blockers, which failed to increase the protein level. Na^+ channel blockers stabilized the short-lived Kir6.2 channel proteins like MG132. There were several SNPs on the lysine residues in the coding region of Kir6.2, suggesting SNPs can affect the extent of channel protein degradation.Conclusion : Expression of short-lived ion channels have been regulated by ubiquitin-proteasome, which can be modulated by Na^+ channel blockers. Based on the SNPs information, a pharmacological therapy to regulate proteasomal degradation of channel protein can be available for treating the patients with chronic atrial fibrillation.

背景：以前，我们报道了负责慢性心房颤动的离子通道已被泛素蛋白蛋白酶体降解，这是抗心律失常剂的修饰。在当前的研究中，我们阐明了其基本机制和应用于对心房颤动的个性化治疗。方法和结果：Na^+通道阻滞剂，但不是Ca^<2+>拮抗剂或K^+通道阻滞剂，或K^+通道阻滞剂阻​​断了20S蛋白酶体的蛋白酶体，以稳定在内，以稳定在内的p53和KCH，IMKK2和KV1.5，IMKK2和KV1.5。转染的COS细胞中的斑块夹技术。为了确认Na^+通道阻滞剂的p53稳定为蛋白酶体抑制作用，我们检查了MDM2（C438A）是否可以阻止药物效应。当与MDM2（C438A）共表达时，即使用MG132处理细胞，p53-FLAG也没有泛素化，并且该药物未能增加p53-FLAG蛋白水平。同样，p53-FLAG也不在用Na^+通道阻滞剂处理的细胞中泛素化，这未能增加蛋白质水平。 Na^+通道阻滞剂稳定了短暂的Kir6.2通道蛋白（如MG132）。在Kir6.2的编码区域中，赖氨酸残基有几个SNP，这表明SNP会影响通道蛋白降解的程度。结论：短寿命的离子通道的表达受到泛素蛋白 - 蛋白酶体的调节，可以通过Na^+通道阻滞剂进行调节。根据SNPS信息，可以使用用于调节通道蛋白蛋白酶体降解的药理疗法可用于治疗慢性心房颤动患者。

项目成果

心筋型ATP感受性Kチャンネルはユビキチン/プロテアソーム系により機能性チャンネル蛋白が規定される。

心脏 ATP 敏感 K 通道的功能通道蛋白由泛素/蛋白酶体系统定义。

• 发表时间: 2004
• 期刊: 心電図 24
• 作者: Kurata Y;et al.;田中宏明
• 通讯作者: 田中宏明

Detection of the novel autoantibody (anti-UACA antibody) in patients with Graves' disease.

• DOI: 10.1016/j.bbrc.2004.06.162
• 发表时间: 2004-08
• 期刊: Biochemical and biophysical research communications
• 影响因子: 3.1
• 作者: T. Ohkura;Shin‐ichi Taniguchi;Kazuhiro Yamada;N. Nishio;T. Okamura;Akio Yoshida;Keiichi Kamijou;S. Fukata;K. Kuma;Y. Inoue;I. Hisatome;S. Senju;Y. Nishimura;C. Shigemasa

Ichida K, et al.: "Clinical and molecular analysis of patients with renal hypouricemia in Japan-influence of URAT1 gene on urinary urate excretion"J Am Soc Nephrol.. 15(1). 164-173 (2004)

Ichida K等人：“日本肾性低尿酸血症患者的临床和分子分析——URAT1基因对尿酸盐排泄的影响”J Am Soc Nephrol.. 15(1)。

State-dependent blocking actions of azimilide dihydrochlo-ride (NE-10064) on human cardiac Na(+) channels.

阿齐利特二盐酸盐 (NE-10064) 对人心脏 Na(+) 通道的状态依赖性阻断作用。

• 发表时间: 2004
• 期刊: Circ J. 68(7)
• 作者: Miake J;et al.
• 通讯作者: et al.

Okamura T, et al.: "Abnormally High Expression of Proteasome Activator-gamma in Thyroid Neoplasm"The Journal of Clinical Endocrinology and Metabolism. 88(3). 1374-1383 (2003)

Okamura T 等人：“甲状腺肿瘤中蛋白酶体激活剂-γ 的异常高表达”《临床内分泌与代谢杂志》。