Biochemical and cell biological characterization of a mammalian septin, Sept 11
Biochemical and cell biological characterization of a mammalian septin, Sept 11
复制标题
DOI：
10.1016/j.febslet.2004.05.030
复制
发表时间：
2004-06-18
期刊：
影响因子：
3.5
通讯作者：
Inagaki, M
中科院分区：
文献类型：
--
作者：
Hanai, N;Nagata, K;Inagaki, M
Septins是一个保守的细胞骨架GTP酶家族，涉及多种细胞功能，如胞质分裂和囊泡运输。在这里，我们报告了一个尚未确定的septin，Sept 11，从猪脑中纯化的septin复合物的鉴定。除白细胞外，在所有受试组织中均检测到转录本。一个Sept 11突变体与明显降低GTdR活性没有形成丝在瞬时表达系统中使用COS 7细胞。通过使用特异性抗体的Western印迹分析，在各种细胞系以及脑组织中检测到Sept 11。用抗Sept 9和抗Sept 11抗体从猪脑中免疫分离的Septin复合物被发现含有基于SDS-PAGE分析的不同的Sept 9同种型，然后通过银染和Western印迹。免疫荧光研究显示细胞类型依赖的细胞内定位的蛋白质; Sept 11共定位主要与微管和肌动蛋白应力纤维在HMEC细胞和REF 52细胞，分别，和他们的丝状分布依赖于细胞骨架结构与蛋白质共定位。Sept 11与HeLa细胞中的应力纤维和微管部分共定位。(C)2004年欧洲生物化学学会联合会。Elsevier B. V.出版，保留所有权利。
Septins are a family of conserved cytoskeletal GTPases implicated in a variety of cellular functions such as cytokinesis and vesicle trafficking. Here, we report identification of an yet uncharacterized septin, Sept11, in septin complexes purified from porcine brain. The transcripts were detected in all tested tissues except leukocytes. A Sept11 mutant with apparently reduced GTPase activity did not form filaments in the transient expression system using COS7 cells. By Western blot analysis using a specific antibody, Sept11 was detected in various cell lines as well as brain tissues. Septin complexes immunoisolated from porcine brain with anti-Sept9 and anti-Sept11 antibodies were found to contain different Sept9 isoforms based on SDS-PAGE analyses followed by silver-staining and Western blotting. Immunofluorescent study revealed cell type-dependent intracellular localization of the protein; Sept11 was colocalized dominantly with microtubules and actin stress fibers in HMEC cells and REF52 cells, respectively, and their filamentous distribution was dependent on the cytoskeleton structures with which the protein is colocalized. Sept11 partially colocalized with stress fibers and microtubules in HeLa cells. (C) 2004 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.