Allsopp T E, Moss D J
M.R.C. Cell Biophysics Unit, Kings College, London, United Kingdom.
J Neurosci. 1989 Jan;9(1):13-24. doi: 10.1523/JNEUROSCI.09-01-00013.1989.
Monoclonal antibody 3D5 recognizes a single component of the neuronal membrane skeleton isolated from the chicken embryo brain. The 3D5 antigen is highly enriched in the CNS, and smaller amounts are found in the PNS. It is also present in non-neural tissues, but this is due to peripheral innervation. The biochemical and molecular properties of the 3D5 antigen are very similar to those of the previously described mammalian protein B-50 (Zwiers et al., 1985)/GAP 43 (Jacobson et al., 1986)/pp46 (Ellis et al., 1985)/F1 (Chan et al., 1986), and include anomalous SDS gel migration, acidic isoelectric point, and extraction from the cytoplasmic side of the plasma membrane only under extremely alkaline conditions. The 3D5 antigen is also developmentally regulated, with maximum expression in brain occurring at E14-E16, after which levels decrease approximately 4-fold in the adult. Immunofluorescence staining of cultured neurons shows that the 3D5 antigen is located in all parts of the cell but is particularly enriched in the growth cone and the growth cone filopodia. As the 3D5 antigen is enriched in the membrane skeleton, we suggest that this protein is involved in an association between the actin-containing cytoskeleton and the plasma membrane.
单克隆抗体3D5识别从鸡胚脑中分离出的神经元膜骨架的单一成分。3D5抗原在中枢神经系统中高度富集,在周围神经系统中含量较少。它也存在于非神经组织中,但这是由于外周神经支配所致。3D5抗原的生化和分子特性与先前描述的哺乳动物蛋白B-50(Zwiers等人,1985年)/GAP 43(Jacobson等人,1986年)/pp46(Ellis等人,1985年)/F1(Chan等人,1986年)非常相似,包括异常的SDS凝胶迁移、酸性等电点,以及仅在极碱性条件下从质膜的细胞质侧提取。3D5抗原也受发育调控,在脑发育过程中,E14 - E16时表达量最高,之后在成体中水平下降约4倍。对培养神经元的免疫荧光染色显示,3D5抗原位于细胞的各个部位,但在生长锥和生长锥丝状伪足中特别富集。由于3D5抗原在膜骨架中富集,我们认为该蛋白参与了含肌动蛋白的细胞骨架与质膜之间的关联。