Oh Ji-eun, Karlmark Karlin Raja, Shin Joo-ho, Pollak Arnold, Freilinger Angelika, Hengstschläger Markus, Lubec Gert
Department of Pediatrics, Medical University of Vienna, Währinger Gürtel 18, A 1090, Vienna, Austria.
Neurochem Res. 2005 Mar;30(3):333-48. doi: 10.1007/s11064-005-2607-2.
No systematic searches for differential expression of signaling proteins (SP) in undifferentiated vs. differentiated cell lineages were published and herein we used protein profiling for this purpose. The NIE-115 cell line was cultivated and an aliquot was differentiated with dimethylsulfoxide (DMSO), that is known to lead to a neuronal phenotype. Cell lysates were prepared, run on two-dimensional gel electrophoresis followed by MALDI-TOF-TOF identification of proteins and maps of identified SPs were generated. Seven SPs were comparable, 27 SPs: GTP-binding/Ras-related proteins, kinases, growth factors, calcium binding proteins, phosphatase-related proteins were observed in differentiated NIE-115 cells and eight SPs of the groups mentioned above were observed in undifferentiated cells only. Switching-on/off of several individual SPs from different signaling cascades during the differentiation process is a key to understand mechanisms involved. The findings reported herein are challenging in vitro and in vivo studies to confirm a functional role for deranged SPs.
此前尚未发表过关于未分化细胞系与分化细胞系中信号蛋白（SP）差异表达的系统性研究，因此我们在此采用蛋白质谱分析来进行此项研究。培养NIE - 115细胞系，取一份样本用二甲基亚砜（DMSO）进行分化处理，已知二甲基亚砜会诱导出神经元表型。制备细胞裂解液，进行二维凝胶电泳，随后通过基质辅助激光解吸电离飞行时间串联质谱（MALDI - TOF - TOF）鉴定蛋白质，并生成已鉴定信号蛋白的图谱。有7种信号蛋白在两者中具有可比性，在分化的NIE - 115细胞中观察到27种信号蛋白：GTP结合/Ras相关蛋白、激酶、生长因子、钙结合蛋白、磷酸酶相关蛋白，而仅在未分化细胞中观察到上述组中的8种信号蛋白。在分化过程中，来自不同信号级联的几种单个信号蛋白的开启/关闭是理解其中涉及机制的关键。本文报道的研究结果对体外和体内研究提出了挑战，以证实紊乱的信号蛋白的功能作用。
