ATP‐dependent copper transport by the Menkes protein in membrane vesicles isolated from cultured Chinese hamster ovary cells

FEBS Letters

Brooks, Hilary¹  Shen, Peiyan¹  Voskoboinik, Ilia¹  Smith, Suzanne²  Camakaris, James¹ 

[1] Department of Genetics, University of Melbourne, Parkville, Vic. 3052, Australia;Radio-Pharmaceutical Division, ANSTO, Lucas Heights Research Laboratories, Lucas Heights, NSW 2234, Australia

关键词: Menkes disease;    Copper;    P-type ATPase;    Copper transport;    AMP-PCP;    adenosine-5′-[β;    γ-methylene]triphosphate;    CHO;    Chinese hamster ovary cell line;    CUR;    copper resistant CHO cell line;    DTT;    dl-dithiothreitol;    MNK;    Menkes protein;    hMNK;    human Menkes protein;    chMNK;    CHO Menkes homologue;    NEM;    N-ethylmaleimide;    PCMB;    p-chloromercuribenzoic acid;   

DOI  :  10.1016/S0014-5793(98)01059-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The Menkes (MNK) protein is a vital component of copper homeostasis in mammalian cells. In this paper we provide the first biochemical evidence that the MNK protein functions as a copper-translocating P-type ATPase in mammalian cells. The enzyme activity in membrane vesicles prepared from Chinese hamster ovary cells overexpressing MNK was ATP-dependent, correlated with the amount of MNK and followed Michaelis-Menten kinetics with respect to copper. The copper transport was observed only under reducing conditions suggesting MNK transports Cu(I). This study opens the way to detailed structure-function studies and assessment of functional MNK derived from patients with Menkes disease.

【 授权许可】

Unknown