Role of Calpain-mediated Cleavage of Calcineurin in Excitotoxic Neurodegeneration

钙蛋白酶介导的钙调磷酸酶裂解在兴奋性神经变性中的作用

• 批准号: 12557116
• 负责人: MATSUI Hideki
• 金额: $ 6.53万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557116/
• 关键词: Calcineurin; Calpain; Calpastatin; Neuronal cell death; Apoptosis; Glutamate; Protein therapy; カルシウムチャネル; 免疫抑制剤; FK506結合蛋白質; シクロスポリン; 膀胱肥大; FK506; 蛋白質セラピー; In vivo Protein Transduction; TAT; Aキナーゼ; 長期増強; PKI

Calcineurin and calpain, a Ca2+/calmodulin-dependent protein phosphatase and a Ca2+-dependent cysteine protease, respectively, mediate neuronal cell death through independent cascades. In this research, we showed that during neuroexcitotoxicity, calcineurin A (CnA) is directly cleaved by calpain in vitro and in vivo, resulting in the enzyme being converted to an active form. Mass spectrometry identified three cleavage sites in CnA, two of which were constitutively active forms. Overexpression of the cleaved CnA induced caspase activity and neuronal cell death. Calpain inhibitors and membrane-permeable calpastatin peptides not only blocked the cleavage of CnA, but also protected against excitotoxic neuronal cell death in vitro and in vivo. These results indicate that CnA is a crucial target for calpain, and the calpain-mediated activation of CnA triggers excitotoxic neurodegeneration. This study established a molecular link between calpain and calcineurin, thereby demonstrating a new mechanism for proteolytical regulation of calcineurin by calpain in response to certain pathological states.

钙调磷酸酶和钙蛋白酶分别是一种 Ca2+/钙调蛋白依赖性蛋白磷酸酶和一种 Ca2+依赖性半胱氨酸蛋白酶，通过独立的级联介导神经元细胞死亡。在这项研究中，我们表明，在神经兴奋毒性过程中，钙调神经磷酸酶 A (CnA) 在体外和体内直接被钙蛋白酶裂解，导致酶转化为活性形式。质谱分析确定了 CnA 中的三个切割位点，其中两个是组成型活性形式。裂解的 CnA 的过度表达诱导 caspase 活性和神经元细胞死亡。钙蛋白酶抑制剂和膜渗透性钙蛋白酶抑制肽不仅能阻断 CnA 的裂解，还能在体外和体内防止兴奋毒性神经元细胞死亡。这些结果表明，CnA 是钙蛋白酶的重要靶标，钙蛋白酶介导的 CnA 激活会引发兴奋性神经变性。这项研究建立了钙蛋白酶和钙调磷酸酶之间的分子联系，从而证明了钙蛋白酶响应某些病理状态对钙调磷酸酶进行蛋白水解调节的新机制。

项目成果

Tomizawa, K., Matsui H.et al.: "Oxytocin improves long-lasting spatial memory in motherhood hippocampus through MAP kinase cascade."Nature Neurosci.. 6(4). 384-390 (2003)

Tomizawa, K., Matsui H.等人：“催产素通过 MAP 激酶级联改善母性海马体的持久空间记忆。”Nature Neurosci.. 6(4)。

富澤一仁, 松下正之, 杉本直巳, 松井秀樹: "化学"遺伝子治療からタンパク質セラピーへ. 5 (2002)

Kazuhito Tomizawa、Masayuki Matsushita、Naomi Sugimoto、Hideki Matsui：“化学”从基因治疗到蛋白质治疗 5 (2002)。

Li, S-T., Kato, K., Tomizawa, K., Matsushita, M., Moriwaki, A., Matsui, H., Mikoshiba, K.: "Calcineurin Plays Different Roles in Group II Metabotropic Glutamate Receptor-and NNIDA Receptor-Dependent Long-Term Depression."J.Neurosci.. 22(12). 5034-5041 (20

Li, S-T.、Kato, K.、Tomizawa, K.、Matsushita, M.、Moriwaki, A.、Matsui, H.、Mikoshiba, K.：“钙调磷酸酶在 II 组代谢型谷氨酸受体和 NNIDA 受体中发挥不同作用

Li, S-T., Kato, K.: "Calcineurin Plays Different Roles in Group II Metabotropic Glutamate Receptor-and NMDA Receptor-Dependent Long-Term Depression"J.Neurosci.. 22(12). 5034-5041 (2002)

Li, S-T., Kato, K.：“钙调神经磷酸酶在 II 组代谢型谷氨酸受体和 NMDA 受体依赖性长期抑郁症中发挥不同作用”J. Neurosci.. 22(12)。

Wu, H-Y., Matsui, H.et al.: "Poly-arginine-fused calpastatin peptide, a livind cell membrane-permeable and specific inhibitor for calpain."Neurosci.Res.. 47. 131-135 (2003)

Wu, H-Y., Matsui, H.et al.：“聚精氨酸融合的钙蛋白酶肽，钙蛋白酶的活细胞膜渗透性和特异性抑制剂。”Neurosci.Res.. 47. 131-135 (2003)