Identification of pleiotrophin in conditioned medium secreted from neural stem cells by SELDI-TOF and SELDI-tandem mass spectrometry
Identification of pleiotrophin in conditioned medium secreted from neural stem cells by SELDI-TOF and SELDI-tandem mass spectrometry
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DOI：
10.1016/j.devbrainres.2004.06.014
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发表时间：
2004-09-17
期刊：
影响因子：
--
通讯作者：
Shiwa, M
中科院分区：
文献类型：
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作者：
Furuta, M;Shiraishi, T;Shiwa, M
神经干细胞（NSC）是具有自我更新活性的多能祖细胞。由于原位 NSC 的命运取决于其含有生长因子和细胞因子的生态位，因此我们采用表面增强激光解吸/电离飞行时间质谱 (SELDI-TOF-MS) 筛选神经干细胞条件培养基中的差异分泌蛋白，并与 NIH3T3 细胞进行比较。通过 SELDI-TOF-MS 和 ProteinChip-tandem MS 系统将仅在神经干细胞条件培养基中检测到的 15.3-kDa 蛋白质确定为多效素 (PTN)。通过一维 SDS 电泳和 Edman 降解分析进一步证实了多效蛋白的鉴定。在神经球中检测到 PTN 及其受体 [受体蛋白酪氨酸磷酸酶 (RPTP) beta/zeta、N-syndecan 和间变性淋巴瘤激酶 (ALK)] 的 mRNA 转录本，表明多效蛋白信号系统存在于神经干细胞中，并参与神经干细胞命运的调节。 (C) 2004 Elsevier B.V. 保留所有权利。
Neural stem cells (NSCs) are multipotential progenitor cells that have self-renewal activity. Since the fates of the NSCs in situ depend on their niche containing growth factors and cytokines, we performed surface enhanced laser desorption/ionization time-of flight mass spectrometry (SELDI-TOF-MS) to screen for differentially secreted proteins in conditioned medium of neural stem cells and compared with that of NIH3T3 cells. A 15.3-kDa protein detected only in the conditioned medium of neural stem cells was determined as pleiotrophin (PTN) by SELDI-TOF-MS and ProteinChip-tandem MS systems. Identification of pleiotrophin was further confirmed by one-dimensional SDS get electrophoresis and Edman degradation analysis. The mRNA transcripts of PTN and its receptors [receptor protein tyrosine phosphatase (RPTP) beta/zeta, N-syndecan and anaplastic lymphoma kinase (ALK)] were detected in neurosphere, suggesting that pleiotrophin signaling systems are present in the neural stem cells and are involved in the modulation of fate of neural stem cells. (C) 2004 Elsevier B.V. All rights reserved.