Oh Ji-Eun, Freilinger Angelika, Gelpi Ellen, Pollak Arnold, Hengstschläger Markus, Lubec Gert
Department of Pediatrics, Medical University of Vienna, Vienna, Austria.
Electrophoresis. 2007 Jun;28(12):2009-17. doi: 10.1002/elps.200600833.
Neuronal differentiation (ND) represents a well-defined phenomenon in biological terms but proteins involved have not been studied systematically. We therefore aimed to study ND by retinoic acid (RA) in a widely used neuroblastoma cell line by comparative proteomics. The ND was induced in the N1E-115 cell line by serum deprivation and RA treatment. Undifferentiated cells and cells undergoing serum deprivation served as controls. Protein extracts were run on 2-DE followed by MALDI-TOF or MALDI-TOF-TOF analysis. Quantification was carried out using specific software and stringent statistical analysis was performed. Tubulin beta 5, cat eye syndrome critical region protein 5 homolog, putative GTP-binding protein PTD004 homolog, and the metabolic proteins glyceraldehyde-3-phosphate dehydrogenase and transketolase were differentially regulated. Differential protein levels of cytoskeleton proteins including tubulins and metabolic proteins have been reported to be regulated by ND. Herein, specific signaling differences as reflected by putative GTP-binding protein PTD004 changes in differentiated cells are shown and a possible role for the Cat eye syndrome critical region protein 5 homolog is proposed. The protein disulfide isomerase associated 3 protein fits the already proposed findings of chaperon regulation by ND. The study forms the molecular basis for further evaluation of the functional roles of the differentially expressed proteins in ND.
神经元分化(ND)在生物学上是一种明确的现象,但尚未对其中涉及的蛋白质进行系统研究。因此,我们旨在通过比较蛋白质组学研究视黄酸(RA)在广泛使用的神经母细胞瘤细胞系中诱导的神经元分化。通过血清剥夺和RA处理在N1E-115细胞系中诱导神经元分化。未分化细胞和经历血清剥夺的细胞用作对照。蛋白质提取物经二维电泳后进行基质辅助激光解吸电离飞行时间(MALDI-TOF)或串联基质辅助激光解吸电离飞行时间(MALDI-TOF-TOF)分析。使用特定软件进行定量,并进行严格的统计分析。微管蛋白β5、猫眼综合征关键区域蛋白5同源物、假定的GTP结合蛋白PTD004同源物以及代谢蛋白甘油醛-3-磷酸脱氢酶和转酮醇酶受到差异调节。据报道,包括微管蛋白在内的细胞骨架蛋白和代谢蛋白的差异蛋白水平受神经元分化调节。在此,展示了分化细胞中假定的GTP结合蛋白PTD004变化所反映的特定信号差异,并提出了猫眼综合征关键区域蛋白5同源物的可能作用。蛋白质二硫键异构酶相关3蛋白符合先前提出的神经元分化对伴侣蛋白调节的研究结果。该研究为进一步评估差异表达蛋白在神经元分化中的功能作用奠定了分子基础。