A NMR study of the interaction of a three-domain construct of ATP7A with copper(I) and copper(I)-HAH1 - The interplay of domains
A NMR study of the interaction of a three-domain construct of ATP7A with copper(I) and copper(I)-HAH1 - The interplay of domains
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DOI：
10.1074/jbc.m506219200
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发表时间：
2005-11-18
影响因子：
4.8
通讯作者：
Rosato, A
中科院分区：
文献类型：
--
作者：
Banci, L;Bertini, I;Rosato, A
ATP 7A是一种P型ATP酶，参与人体铜（I）稳态。它具有一个长的N-末端尾突出到胞质溶胶，并含有六个铜（I）结合域，这是单独折叠，并能够结合一个铜（I）离子。ATP 7A从可溶性蛋白质金属伴侣HAH 1接收铜。这六个可溶性结构域的确切作用和相互作用仍然非常不清楚，因为已经广泛证明它们在体内铜（I）转运方面是非常冗余的。在目前的工作中，三域（第四至第六，MNK 456）的结构已在溶液中进行了研究，通过NMR，在存在和不存在的铜（I）。此外，还研究了MNK 456与Cu（I）-HAH 1的相互作用。据推测，第四个结构域是与合作伙伴进行初始相互作用的优先位点。一个显着的依赖性的整体域动态的metaphoric状态和HAH 1的存在下进行观察。这种依赖性可能构成触发铜（I）易位和/或ATP 7A从trans-Golgi网络重新定位到质膜的分子机制。
ATP7A is a P-type ATPase involved in copper(I) homeostasis in humans. It possesses a long N-terminal tail protruding into the cytosol and containing six copper(I)-binding domains, which are individually folded and capable of binding one copper(I) ion. ATP7A receives copper from a soluble protein, the metallochaperone HAH1. The exact role and interplay of the six soluble domains is still quite unclear, as it has been extensively demonstrated that they are strongly redundant with respect to copper(I) transport in vivo. In the present work, a three-domain (fourth to sixth, MNK456) construct has been investigated in solution by NMR, in the absence and presence of copper(I). In addition, the interaction of MNK456 with copper(I)-HAH1 has been studied. It is proposed that the fourth domain is the preferential site for the initial interaction with the partner. A significant dependence of the overall domain dynamics on the metallation state and on the presence of HAH1 is observed. This dependence could constitute the molecular mechanism to trigger copper(I) translocation and/or ATP7A relocalization from the trans-Golgi network to the plasmatic membrane.