Wakatsuki S, Ho S C, Arioka M, Yamasaki M, Kitamoto K
Department of Biotechnology, The University of Tokyo.
J Biochem. 1997 Dec;122(6):1146-52. doi: 10.1093/oxfordjournals.jbchem.a021874.
By screening with monoclonal antibodies (mAbs) raised against growth cone membrane fraction from fetal porcine brains, we have identified a 230 kDa antigen, termed p230. Western blot analysis of extracts from various tissues demonstrated that p230 is specifically expressed in brains, in which its expression is temporally restricted; it was especially prominent in the embryonic and the early postnatal stage, and decreased to subdetectable levels in the adult brain. Further characterization of p230 revealed that it is a peripherally-membrane associated, cell surface protein produced by astrocytes. Neurite outgrowth of E18 rat cerebral cortex neurons cultured on a monolayer of astrocytes was significantly reduced in the presence of anti-p230 polyclonal antibody. Partial amino acid sequences of p230 purified from fetal porcine brains were highly homologous to an extracellular matrix protein, tenascin-C. These lines of evidence suggest that p230, a tenascin-C-like molecule present in fetal porcine brains, plays important roles during early brain development, particularly in growth cone guidance.
通过用针对胎猪脑生长锥膜组分产生的单克隆抗体(mAb)进行筛选,我们鉴定出一种230 kDa的抗原,称为p230。对来自各种组织的提取物进行的蛋白质印迹分析表明,p230在脑中特异性表达,其表达在时间上受到限制;它在胚胎期和出生后早期尤为突出,在成体脑中降至检测不到的水平。对p230的进一步表征表明,它是一种由星形胶质细胞产生的外周膜相关细胞表面蛋白。在抗p230多克隆抗体存在的情况下,培养在星形胶质细胞单层上的E18大鼠大脑皮质神经元的神经突生长显著减少。从胎猪脑中纯化的p230的部分氨基酸序列与细胞外基质蛋白腱生蛋白-C高度同源。这些证据表明,p230是胎猪脑中存在的一种腱生蛋白-C样分子,在早期脑发育过程中起重要作用,特别是在生长锥导向方面。