Identification and characterization of a novel Cut family cDNA that encodes human copper transporter protein CutC.
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.
Li
Jixi
J
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, People's Republic of China.
Ji
Chaoneng
C
Chen
Jinzhong
J
Yang
Zhenxing
Z
Wang
Yijing
Y
Fei
Xiangwei
X
Zheng
Mei
M
Gu
Xing
X
Wen
Ge
G
Xie
Yi
Y
Mao
Yumin
Y
D000595:Amino Acid Sequence; D027682:Cation Transport Proteins / Q000032:analysis* / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D019295:Computational Biology; D003300:Copper / Q000378:metabolism; D000079922:Copper Transport Proteins; D018076:DNA, Complementary / Q000737:chemistry; D006801:Humans; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D014018:Tissue Distribution
D027682:Cation Transport Proteins; D003300:Copper; D000079922:Copper Transport Proteins; D018076:DNA, Complementary; C505249:CUTC protein, human