Johansen K M, Johansen J
Department of Zoology and Genetics, Iowa State University, Ames 50011, USA.
J Neurobiol. 1995 Jun;27(2):227-39. doi: 10.1002/neu.480270209.
The lan 3-8 monoclonal antibody recognizes a 63 kD antigen that is associated with the cytoskeleton in leech neurons. We have used this antibody to clone a novel invertebrate neuronal intermediate filament protein, filarin, by screening an expression vector library. A full-length clone of 2.2 kb identified by the antibody was isolated and sequenced. The protein contains a coiled-coil rod domain typical of the superfamily of intermediate filament proteins flanked by unique N- and C-terminal domains. The highest homology of filarin is to the alternatively spliced squid brain intermediate filament protein (Szaro et al., 1991, J. Biol. Chem. 266:15035-15041), the only other invertebrate neuronal intermediate filament in the data bank. However, apart from extensive homology in the two end regions of the rod domain, the similarity of the two proteins is limited to the general coiled-coil structure of intermediate filaments. Thus, filarin may represent a novel type of invertebrate neuronal intermediate filament protein. Filarin contains the extra 6 heptads characteristic of lamins and of all cytoplasmic invertebrate intermediate filaments analyzed so far. By Northern analysis, it appears that filarin is not alternatively spliced, since only a single transcript of 2.2 kb is recognized by the clone. Using the lan 3-8 antibody to follow its developmental expression, we found that filarin is present in all known neurons in the central and peripheral nervous system.
lan 3 - 8单克隆抗体识别一种63 kD的抗原,该抗原与水蛭神经元中的细胞骨架相关。我们利用这种抗体通过筛选表达载体文库克隆了一种新型无脊椎动物神经元中间丝蛋白——丝连蛋白。分离并测序了由该抗体鉴定出的一个2.2 kb的全长克隆。该蛋白包含一个中间丝蛋白超家族典型的卷曲螺旋杆状结构域,两侧是独特的N端和C端结构域。丝连蛋白与选择性剪接的鱿鱼脑中间丝蛋白(Szaro等人,1991年,《生物化学杂志》266:15035 - 15041)同源性最高,数据库中唯一的另一种无脊椎动物神经元中间丝蛋白。然而,除了杆状结构域两端区域有广泛的同源性外,这两种蛋白质的相似性仅限于中间丝的一般卷曲螺旋结构。因此,丝连蛋白可能代表一种新型的无脊椎动物神经元中间丝蛋白。丝连蛋白含有核纤层蛋白以及迄今为止分析的所有细胞质无脊椎动物中间丝特有的额外6个七肽重复序列。通过Northern分析,似乎丝连蛋白没有选择性剪接,因为该克隆只识别到一个2.2 kb的单一转录本。利用lan 3 - 8抗体追踪其发育表达情况,我们发现丝连蛋白存在于中枢和外周神经系统的所有已知神经元中。