Meyerson G, Pfenninger K H, Påhlman S
Department of Pathology, University of Uppsala, University Hospital, Sweden.
J Cell Sci. 1992 Sep;103 ( Pt 1):233-43. doi: 10.1242/jcs.103.1.233.
Nerve growth cones of primary neurons are highly enriched in the proto-oncogene product pp60c-src. In order to investigate this molecule further in growing neuronal cells, growth cone and cell body fractions were prepared from human SH-SY5Y neuroblastoma cells differentiated neuronally in vitro under the influence of phorbol ester. The fractions were characterized ultrastructurally and by biochemical criteria. The neuronal (pp60c-srcN) and the fibroblastic (pp60c-src) forms of pp60src are slightly enriched and activated in the growth cones relative to the perikarya. Immunoprecipitates of pp60src from differentiated SH-SY5Y growth cones contain at least four phosphoproteins in addition to pp60src. One of these, pp38, migrates as a 100-140 kDa complex with pp60src under non-reducing conditions of gel electrophoresis. The pp38/pp60src complex is not easily detected in non-differentiated SH-SY5Y cells or perikarya of differentiated SH-SY5Y cells, but it is highly enriched in the growth cone preparation. These data suggest that growth-cone pp60src exists in a disulfide-linked oligomeric complex. The complex appears to be assembled only in the cell periphery and may be dependent upon neuronal differentiation.
原代神经元的神经生长锥中高度富含原癌基因产物pp60c-src。为了在生长中的神经元细胞中进一步研究该分子,在佛波酯的影响下,从体外神经元分化的人SH-SY5Y神经母细胞瘤细胞制备生长锥和细胞体部分。通过超微结构和生化标准对这些部分进行表征。相对于核周体,pp60src的神经元形式(pp60c-srcN)和成纤维细胞形式(pp60c-src)在生长锥中略有富集并被激活。来自分化的SH-SY5Y生长锥的pp60src免疫沉淀物除了pp60src外还包含至少四种磷蛋白。其中之一pp38,在凝胶电泳的非还原条件下与pp60src以100 - 140 kDa复合物的形式迁移。pp38/pp60src复合物在未分化的SH-SY5Y细胞或分化的SH-SY5Y细胞的核周体中不易检测到,但在生长锥制备物中高度富集。这些数据表明生长锥pp60src以二硫键连接的寡聚复合物形式存在。该复合物似乎仅在细胞周边组装,并且可能依赖于神经元分化。