Oligomeric structure of bAE3 protein.
Oligomeric structure of bAE3 protein.
复制标题
bAE3 蛋白的寡聚结构。
DOI：
10.1080/713803739
复制
发表时间：
2000
期刊：
影响因子：
--
通讯作者：
Kurtz,I
中科院分区：
文献类型：
--
作者：
Pushkin,AV;Tsuprun,VL;Abuladze,NK;Newman,D;Kurtz,I
通过差速离心和免疫亲和层析，从兔肾中纯化出“脑”形式的阴离子交换蛋白3 （bAE3）。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和Western blotting检测到一个»165 kDa的单蛋白条带。纯化的bAE3在氧化交联后发现了单体、二聚体（主要成分）和更高的低聚形式（显然是四聚体）。通过蔗糖梯度离心分离得到了最大形式的bAE3二聚体和单体，并通过透射电镜对其进行了研究，证实其为四聚体结构。检测到两种主要类型的bAE3图像，圆形（»11-14 nm）和方形（»12 - 12 nm）。图像分析显示，圆形和方形图像均具有四倍旋转对称性，表明bAE3由4个亚基的倍数组成。我们得出结论，在Triton X-100溶液中，bAE3主要是二聚体和四聚体的混合物，单体数量较少。
The “brain” form of the anion exchanger protein 3 (bAE3) has been puri ed to homogeneity from the rabbit kidney by differential centrifugation and immunoaf nity chromatography. A single protein band of» 165 kDa was detected by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) and Western blotting. Monomers, dimers (a major component), and a higher oligomeric form (apparently tetramers) were found after oxidative cross-linking of puri ed bAE3. The largest form of bAE3 was separated from dimers and monomers by sucrose gradient centrifugation and was studied by transmission electron microscopy to conrm a tetrameric structure. Two main types of bAE3 images were detected, round (» 11–14 nm) and square-shaped (» 12£ 12 nm). Image analysis revealed fourfold rotational symmetry of both the round and square-shaped images, indicating that bAE3 consists of multiples of 4 subunits. We conclude that bAE3 in Triton X-100 solution is predominantly a mixture of dimers and tetramers with a smaller amount of monomers.