Li Jixi, Ji Chaoneng, Chen Jinzhong, Yang Zhenxing, Wang Yijing, Fei Xiangwei, Zheng Mei, Gu Xing, Wen Ge, Xie Yi, Mao Yumin
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, People's Republic of China.
Biochem Biophys Res Commun. 2005 Nov 11;337(1):179-83. doi: 10.1016/j.bbrc.2005.09.029.
Copper is an essential heavy metal trace element that plays important roles in cell physiology. The Cut family was associated with the copper homeostasis and involved in several important metabolisms, such as uptake, storage, delivery, and efflux of copper. In this study, a novel Cut family cDNA was isolated from the human fetal brain library, which encodes a 273 amino acid protein with a molecular mass of about 29.3 kDa and a calculated pI of 8.17. It was named hCutC (human copper transporter protein CutC). The ORF of hCutC gene was cloned into pQE30 vector and expressed in Escherichia coli M15. The secreted hCutC protein was purified to a homogenicity of 95% by using the Ni-NTA affinity chromatography. RT-PCR analysis showed that the hCutC gene expressed extensively in human tissues. Subcellular location analysis of hCutC-EGFP fusion protein revealed that hCutC was distributed to cytoplasm of COS-7 cells, and both cytoplasm and nucleus of AD293 cells. The results suggest that hCutC may be one shuttle protein and play important roles in intracellular copper trafficking.
铜是一种必需的重金属微量元素，在细胞生理学中发挥着重要作用。Cut家族与铜稳态相关，并参与多种重要代谢过程，如铜的摄取、储存、运输和外排。在本研究中，从人胎脑文库中分离出一个新的Cut家族cDNA，它编码一个由273个氨基酸组成的蛋白质，分子量约为29.3 kDa，计算所得的等电点为8.17。它被命名为hCutC（人铜转运蛋白CutC）。将hCutC基因的开放阅读框克隆到pQE30载体中，并在大肠杆菌M15中表达。通过镍-氮三乙酸亲和层析将分泌的hCutC蛋白纯化至95%的纯度。逆转录-聚合酶链反应分析表明，hCutC基因在人体组织中广泛表达。hCutC-增强绿色荧光蛋白融合蛋白的亚细胞定位分析显示，hCutC分布于COS-7细胞的细胞质以及AD293细胞的细胞质和细胞核中。结果表明，hCutC可能是一种穿梭蛋白，在细胞内铜转运中发挥重要作用。
