Functional proteomics analysis of non-vesicular release mechanisms in stress exposed neuron-role of stress-induced non-vesicular release Prion, NDI and FGF-1 in neuron

应激暴露神经元非囊泡释放机制的功能蛋白质组学分析应激诱导非囊泡释放朊病毒、NDI和FGF-1在神经元中的作用

• 批准号: 15390028
• 负责人: UEDA Hiroshi
• 金额: $ 9.86万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 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-15390028/
• 关键词: Stress; non-vesicular release; S100A13; FGF-1; serum starvation; Ca^<2+>; STI; NDI; 神経細胞死; ネクローシス; 分泌; 核移行; 低酸素; S100

The purpose of this research is to demonstrate the stress-induced mechanism of non-vesicular release FGF-1, NDI and STI, as prion (PrP^c) associated protein using the proteomics analyses. We attempted to characterize the non-vesicular mode of FGF-1 and NDI release in the analyses using immunocytochemistry and immunoblot of conditioned medium (CM) from astrocytes. FGF-1 was completely released from astrocytes upon serum-deprivation stress in a Brefeldin A-insensitive manner. In the immunoprecipitation study using anti-FGF-1 IgG, S100A13 was identified to be the major protein co-eluted with FGF-1. The interaction between GST-FGF-1 and Strep-tag II S100A13 was found to be Ca^<2+>-sensitive, and to require the C-terminal 11 amino acid peptide sequence of S100A13. The overexpression of D88-98 mutant of S100A13 selectively inhibited the serum-deprivation stress-induced release of FGF-1, but not the release of S100A13 mutant from C6 glioma cells. However, amlexanox, anti-allergic drug whose t … More arget is S100A13, completely inhibited the stress-induced release of FGF-1 as well as S100A13. The stress-induced release of both proteins was also abolished by BAPTA-AM, an intracellular Ca^<2+> chelating agent. The serum-deprivation caused Ca^<2+> spikes in v-conotoxin GVIA and thapsigargin-sensitive manner. These same results observed NDI protein experiments. These results suggest that S100A13 is a cargo molecule for the serum-deprivation stress-induced non-vesicular release of FGF-1 and NDI, and that its driving force of protein-protein interaction and release is possibly mediated by Ca^<2+>-induced Ca^<2+> release (CICR) coupled to N-type Ca^<2+> channel activity. On the other hand, STI have neuronal survival activity. STI inhibitor, as a PrP^c c terminal peptide (113-132), treated neuron was enhanced decrease in survival activity under the oxygen-glucose deprivation. Moreover, STI inhibitor induced neuronal cell death under the serum containing normal condition. These results suggested that PrP^c have increase in neuronal survival activity under the normal condition. Less

本研究的目的是利用蛋白质组学技术研究应激诱导的FGF-1、NDI和STI作为朊病毒（PrP^c）相关蛋白的非囊泡释放机制。我们试图用免疫细胞化学和免疫印迹分析星形胶质细胞的条件培养基（CM），以表征FGF-1和NDI释放的非囊泡模式。FGF-1完全释放星形胶质细胞后血清剥夺应力在布雷菲德菌素A不敏感的方式。在使用抗FGF-1 IgG的免疫沉淀研究中，S100 A13被鉴定为与FGF-1共洗脱的主要蛋白质。发现GST-FGF-1和Strep标签II S100 A13之间的相互作用是Ca^<2+>敏感的，并且需要S100 A13的C末端11个氨基酸的肽序列。过表达S100 A13突变体D88-98选择性抑制血清剥夺应激诱导的C6胶质瘤细胞FGF-1的释放，但不抑制S100 A13突变体的释放。然而，抗过敏药物氨来沙诺， ...更多信息 靶点为S100 A13，与S100 A13一样完全抑制应激诱导的FGF-1释放。BAPTA-AM（一种细胞内Ca^<2+>螯合剂）也能消除应激诱导的两种蛋白质的释放。血清剥夺引起v-芋螺毒素GVIA和毒胡萝卜素敏感的Ca^2+峰。这些相同的结果观察到NDI蛋白实验。这些结果表明，S100 A13是血清剥夺应激诱导的FGF-1和NDI的非囊泡释放的货物分子，并且其蛋白质-蛋白质相互作用和释放的驱动力可能是由Ca^2+诱导的Ca^2+释放（CICR）与N型Ca^2+通道活性偶联介导的。另一方面，STI具有神经元存活活性。STI抑制剂作为PrP^c c末端肽（113-132），可使缺氧缺糖神经元存活活性明显降低。此外，STI抑制剂在含血清的正常条件下诱导神经元细胞死亡。这些结果表明，在正常情况下，PrP^c具有增强神经元存活活性的作用。少

项目成果

Fujita, R., Ueda, H: "Protein kinase C-mediated cell death mode switch induced by high glucose"Cell Death Differentiation. 10. 1336-1347 (2003)

Fujita, R., Ueda, H：“高葡萄糖诱导的蛋白激酶 C 介导的细胞死亡模式转换”细胞死亡分化。

Voltage-dependent N-type Ca2+ channel activity regulates the interaction between FGF-1 and S100A13 for stress-induced non-vesicular release

• DOI: 10.1007/s10571-006-9016-1
• 发表时间: 2006-05-01
• 期刊: CELLULAR AND MOLECULAR NEUROBIOLOGY
• 影响因子: 4
• 作者: Matsunaga, Hayato;Ueda, Hiroshi
• 通讯作者: Ueda, Hiroshi

Neuroprotective drug(5), Neuronal protection by neuronal cell death mode switch and glial activation.

神经保护药物(5)，通过神经元细胞死亡模式切换和神经胶质激活来保护神经元。

• 发表时间: 2006
• 期刊: Clinical Neuroscience 24
• 作者: H.Ueda

Evidence for serum-deprivation-induced co-release of FGF-1 and S100A13 from astrocytes

• DOI: 10.1016/j.neuint.2006.01.017
• 发表时间: 2006-08-01
• 期刊: NEUROCHEMISTRY INTERNATIONAL
• 影响因子: 4.2
• 作者: Matsunaga, Hayato;Ueda, Hiroshi
• 通讯作者: Ueda, Hiroshi

神経保護作用薬(5)神経細胞死モードスイッチによる保護機構とグリア細胞応答

神经保护药物（五）神经元死亡模式切换的保护机制及胶质细胞反应

• 发表时间: 2006
• 期刊: Clinical Neuroscience 24
• 作者: Nishida;M.;Tanabe;S.;Maruyma;Y.;Mangmool;S.;Urayama;K.;Nagamatsu;Y.;Takagahara;S.;Turner;J.H.;Kozasa;T.;Kobayashi;H.;Sato;Y.;Kawanishi;T.;Inoue;R.;Nagao;T.;Kurose;H.;Watanabe S et al.;H.Ueda
• 通讯作者: H.Ueda