Oh J-E, Karlmark Raja K, Shin J-H, Pollak A, Hengstschläger M, Lubec G
Department of Pediatrics, Medical University of Vienna, Vienna, Austria.
Amino Acids. 2006 Oct;31(3):289-98. doi: 10.1007/s00726-005-0256-z. Epub 2006 Mar 20.
No systematic approach to detect expression of differentiation-related elements was published so far. The undifferentiated N1E-115 neuroblastoma cell line was switched into a neuronal phenotype by DMSO treatment and used for proteomic experiments. We used two-dimensional gel electrophoresis followed by unambiguous mass spectrometrical identification of proteins to generate a map of cytoskeleton proteins (CPs), i.e., to search for differentiation-related structures. Alpha-actin, actin-like protein 6A, gamma-tubulin complex component 2, tubulin alpha 3/alpha 7, CLIP associating protein 2, B4 integrin interactor homolog were detectable in the undifferentiated cell line exclusively and neuron-specific CPs drebrin and presynaptic density protein 95, actin-related protein 2/3, alpha and beta-centractin, PDZ-domain actin binding protein, actinin alpha 1, profilin II, ezrin, coactosin-like protein, transgelin 2, myosin light polypeptide 6, tubulin alpha 2, 6 and 7, beta tubulin (94% similar with tubulin beta-2), tubulin beta 3, tubulin tyrosine ligase-like protein 1, lamin B1 and keratin 20 were observed in the differentiated cell line only. We herein identified differentiation-related expressional patterns thus providing new evidence for the role of CPs in the process of neuronal differentiation.
迄今为止，尚未发表检测分化相关元件表达的系统方法。未分化的N1E-115神经母细胞瘤细胞系经二甲基亚砜（DMSO）处理后转变为神经元表型，并用于蛋白质组学实验。我们采用二维凝胶电泳，随后通过明确的蛋白质质谱鉴定来生成细胞骨架蛋白（CPs）图谱，即寻找与分化相关的结构。α-肌动蛋白、肌动蛋白样蛋白6A、γ-微管蛋白复合体组分2、微管蛋白α3/α7、CLIP相关蛋白2、B4整合素相互作用蛋白同源物仅在未分化细胞系中可检测到，而神经元特异性CPs drebrin和突触前密度蛋白95、肌动蛋白相关蛋白2/3、α和β-中心肌动蛋白、PDZ结构域肌动蛋白结合蛋白、肌动蛋白α1、丝切蛋白II、埃兹蛋白、类辅肌动蛋白、转胶蛋白2、肌球蛋白轻链多肽6、微管蛋白α2、6和7、β-微管蛋白（与β-微管蛋白2相似度为94%）、微管蛋白β3、微管蛋白酪氨酸连接酶样蛋白1、核纤层蛋白B1和角蛋白20仅在分化细胞系中观察到。我们在此鉴定出与分化相关的表达模式，从而为CPs在神经元分化过程中的作用提供了新证据。
