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AtKP1是一种类驱动蛋白,主要定位于拟南芥的线粒体中。

AtKP1, a kinesin-like protein, mainly localizes to mitochondria in Arabidopsis thaliana.

作者信息

Ni Cheng Zhi, Wang Hai Qing, Xu Tao, Qu Zhe, Liu Guo Qin

机构信息

State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing.

出版信息

Cell Res. 2005 Sep;15(9):725-33. doi: 10.1038/sj.cr.7290342.

Abstract

Kinesins and kinesin-like proteins (KLPs) constitute a large family of microtubule-based motors that play important roles in many fundamental cellular and developmental processes. To date, a number of kinesins or KLPs have been identified in plants including Arabidopsis thaliana. Here, a polyclonal antibody against AtKP1 (kinesin-like protein 1 in A. thaliana) was raised by injection the expressed AtKP1 specific C-terminal polypeptides in rabbits, and immunoblot analysis was conducted with the affinity-purified anti-AtKP1 antibody. The results indicated that this antibody recognized the AtKP1 fusion proteins expressed in E. coli and proteins of ~125 kDa in the soluble fractions of Arabidopsis extracts. The molecular weight was consistent with the calculated molecular weight based on deduced amino acids sequence of AtKP1. To acquire the subcellular localization of the protein, AtKP1 in Arabidopsis root cells was observed by indirect immunofluorescence microscopy. AtKP1 was localized to particle-like organelles in interphase or dividing cells, but not to mitotic microtubule arrays. Relatively more AtKP1 was found in isolated mitochondria fraction on immunoblot of the subcellular fractions. The AtKP1 protein could not be released following a 0.6 M KI washing, indicating that AtKP1 is tightly bind to mitochondria and might function associated with this kind of organelles.

摘要

驱动蛋白和驱动蛋白样蛋白(KLPs)构成了一个基于微管的马达蛋白大家族,在许多基本的细胞和发育过程中发挥着重要作用。迄今为止,在包括拟南芥在内的植物中已鉴定出许多驱动蛋白或KLPs。在此,通过将表达的拟南芥AtKP1特异性C末端多肽注射到兔子体内,制备了针对AtKP1(拟南芥驱动蛋白样蛋白1)的多克隆抗体,并用亲和纯化的抗AtKP1抗体进行了免疫印迹分析。结果表明,该抗体识别在大肠杆菌中表达的AtKP1融合蛋白以及拟南芥提取物可溶部分中约125 kDa的蛋白。该分子量与基于AtKP1推导氨基酸序列计算出的分子量一致。为了获得该蛋白的亚细胞定位,通过间接免疫荧光显微镜观察了拟南芥根细胞中的AtKP1。AtKP1定位于间期或分裂细胞中的颗粒状细胞器,但不定位于有丝分裂微管阵列。在亚细胞组分的免疫印迹中,在分离的线粒体组分中发现相对较多的AtKP1。用0.6 M KI洗涤后,AtKP1蛋白不能被释放,这表明AtKP1与线粒体紧密结合,可能与这种细胞器相关发挥作用。

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