Voltage-dependent N-type Ca2+ channel activity regulates the interaction between FGF-1 and S100A13 for stress-induced non-vesicular release
Voltage-dependent N-type Ca2+ channel activity regulates the interaction between FGF-1 and S100A13 for stress-induced non-vesicular release
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DOI：
10.1007/s10571-006-9016-1
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发表时间：
2006-05-01
影响因子：
4
通讯作者：
Ueda, Hiroshi
中科院分区：
文献类型：
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作者：
Matsunaga, Hayato;Ueda, Hiroshi
1.研究了Ca ~（2+）对NG 108 -15细胞中FGF-1与S100 A13相互作用的调节作用。当从培养物中去除B27补充物时，细胞水平降低，而它们的释放在3 h内显著增加。抗过敏药氨来沙诺可消除这些应激引起的变化.这些释放显着抑制EGTA或BAPTA-AM，细胞或细胞外的钙离子螯合剂，分别。ω-芋螺毒素GVIA，一种N-型钙通道阻滞剂的加入引起的释放的完全抑制，同时增加了这两种蛋白质的胞质水平。而非N型钙通道阻断剂ω-芋螺毒素MVIIC则无效.在用Venus-FGF-1和CFP-S100 A13转染的NG 108 -15细胞中，剥夺顺应性应激引起几个尖峰型荧光共振能量转移（FRET）信号，表明显示相互作用的两种蛋白质将立即释放。通过添加ω-芋螺毒素GVIA完全消除这些尖峰。然而，氨来赞的加入导致钟形FRET信号没有尖峰.因此，这表明FGF-1和S100 A13之间的相互作用，负责应力诱导的非囊泡释放是依赖于通过N型钙离子通道的钙离子内流。
1. The Ca2+-mediated regulation of interaction between FGF-1 and S100A13 in NG108-15 cells was studied. When the stress by depriving B27 supplement from the culture was given, cellular levels of both proteins were decreased, while their releases were significantly increased within 3 h. These stress-induced changes were all abolished by amlexanox, an anti-allergic drug.2. These releases were significantly inhibited by the addition of EGTA or BAPTA-AM, cellular or extracellular Ca2+-chelating agent, respectively. The addition of omega-conotoxin GVIA, a N-type Ca2+-channel blocker caused a complete inhibition of the release, while increased the cytosolic levels of both proteins. However, omega-conotoxin MVIIC, the non-N-type Ca2+-channel blocker was ineffective.3. In NG108-15 cells, which had been transfected with Venus-FGF-1 and CFP-S100A13, the supplement-deprivation stress caused several spike-type fluorescence resonance energy transfer (FRET) signals, suggesting that both proteins showing interaction would be immediately released. These spikes were completely abolished by the addition of omega-conotoxin GVIA. However, the addition of amlexanox caused bell-shaped FRET signals without spikes.4. Thus, it is suggested that the interaction between FGF-1 and S100A13 responsible for stress-induced non-vesicular release is dependent of Ca2+-influx through N-type Ca2+-channels.