Ho S C, Wakatsuki S, Arioka M, Yamasaki M, Kitamoto K
Department of Biotechnology, The University of Tokyo.
J Biochem. 1998 Feb;123(2):332-8. doi: 10.1093/oxfordjournals.jbchem.a021941.
To identify and analyze the function of proteins expressed in the growth cones, we have screened monoclonal antibodies raised against the preparation of the growth cone particles derived from fetal porcine brains and found a novel neuronal antigen, termed NP-190. Biochemical characterization of NP-190 demonstrated that it was an integral membrane protein with an apparent molecular weight of 190 kDa and that it was mainly expressed in fetal brains. Homologous antigens with molecular weights of 200 and 170 kDa were also identified in the fetal brain extracts of chickens and rats, respectively. Immunoblot experiments of brain extracts from chickens and rats in various stages of development indicated that the expression of NP-190 homologs was developmentally regulated; it began to appear and increased in the embryonic stage, then decreased to very low level in the adult brains. Immunostaining of cultured primary of neurons from the embryonic day 18 rat cerebral cortex demonstrated that rat NP-190 homolog localized in the cell bodies, axons and growth cones, but not in dendrites. Partial amino acid sequence analysis of affinity-purified NP-190 from fetal porcine brains demonstrated that it was a novel protein. These results suggest that NP-190 plays a distinct role in brain development.
为了鉴定和分析生长锥中表达的蛋白质的功能,我们筛选了针对源自胎猪脑的生长锥颗粒制备物产生的单克隆抗体,并发现了一种新的神经元抗原,称为NP-190。NP-190的生化特性表明,它是一种表观分子量为190 kDa的整合膜蛋白,主要在胎脑中表达。在鸡和大鼠的胎脑提取物中也分别鉴定出分子量为200和170 kDa的同源抗原。对鸡和大鼠不同发育阶段脑提取物的免疫印迹实验表明,NP-190同源物的表达受发育调控;它在胚胎期开始出现并增加,然后在成年脑中降至极低水平。对来自胚胎第18天大鼠大脑皮层的原代培养神经元进行免疫染色表明,大鼠NP-190同源物定位于细胞体、轴突和生长锥,但不在树突中。对从胎猪脑中亲和纯化的NP-190进行的部分氨基酸序列分析表明,它是一种新蛋白。这些结果表明,NP-190在脑发育中起独特作用。