Evidence for serum-deprivation-induced co-release of FGF-1 and S100A13 from astrocytes
Evidence for serum-deprivation-induced co-release of FGF-1 and S100A13 from astrocytes
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DOI：
10.1016/j.neuint.2006.01.017
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发表时间：
2006-08-01
影响因子：
4.2
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
由于成纤维细胞生长因子-1缺乏内质网-高尔基体途径所必需的传统氨基端信号肽，其释放方式尚不完全清楚。我们试图在免疫细胞化学和星形胶质细胞条件培养液(CM)的免疫印迹分析中表征非经典(非囊泡)的成纤维细胞生长因子-1的释放模式。星形胶质细胞在血清剥夺应激条件下可完全释放成纤维细胞生长因子-1，且Brefeldin A不敏感。在抗成纤维细胞生长因子-1抗体的免疫沉淀研究中，S100A13被确定为与成纤维细胞生长因子-1共洗脱的主要蛋白质。GST-FGF1与Strep-Tag II-S S100A13之间的相互作用是钙敏感的，需要S100A13的C-末端11个氨基酸序列。S100A13的Delta 88-98突变体的过表达选择性地抑制血清剥夺应激诱导的C6胶质瘤细胞释放成纤维细胞生长因子-1，但不抑制突变体S100A13的释放。而以S100A13为靶点的抗过敏药物氨来昔诺可完全抑制应激诱导的成纤维细胞生长因子-1和S100A13的释放。应激诱导的这两种蛋白的释放也被细胞内的钙离子螯合剂BAPTA-AM所消除。血清剥夺引起Omega-conooxinGVIA和thapsigargin敏感的钙离子峰值。这些结果表明，S100A13是去血清应激诱导的成纤维细胞生长因子-1非经典释放的货运分子，其蛋白-蛋白质相互作用和释放的驱动力可能是通过钙离子诱导的钙释放(CICR)与N型钙通道活动相耦合。(C)2006爱思唯尔有限公司。保留所有权利。
Since fibroblast growth factor (FGF)-1 lacks conventional amino-terminal signal peptide essential for endoplasmic reticulum (ER)-Golgi pathway, the mode of release of this polypeptide remains to be fully understood. We attempted to characterize the non-classical (non-vesicular) mode of FGF-1 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-S S100A13 was found to be Ca2+ sensitive, and to require the C-terminal 11 amino acid peptide sequence of S100A13. The overexpression of Delta 88-98 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 target 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 Ca2+ chelating agent. The serum-deprivation caused Ca2+ spikes in omega-conotoxin GVIA and thapsigargin-sensitive manner. All these results suggest that S100A13 is a cargo molecule for the serum-deprivation stress-induced non-classical release of FGF-1, and that its driving force of protein-protein interaction and release is possibly mediated by Ca2+-induced Ca2+ release (CICR) coupled to N-type Ca2+ channel activity. (c) 2006 Elsevier Ltd. All rights reserved.