通过预稳态电学测量证明了 ATP7B Cu+-ATPase 中的 ATP 依赖性电荷移动。
ATP dependent charge movement in ATP7B Cu+-ATPase is demonstrated by pre-steady state electrical measurements.
机构信息
Department of Chemistry Ugo Schiff, University of Florence, 50019 Sesto Fiorentino, Italy.
出版信息
FEBS Lett. 2010 Nov 19;584(22):4619-22. doi: 10.1016/j.febslet.2010.10.029. Epub 2010 Oct 21.
Abstract
ATP7B is a copper dependent P-type ATPase, required for copper homeostasis. Taking advantage of high yield heterologous expression of recombinant protein, we investigated charge transfer in ATP7B. We detected charge displacement within a single catalytic cycle upon ATP addition and formation of phosphoenzyme intermediate. We attribute this charge displacement to movement of bound copper within ATP7B. Based on specific mutations, we demonstrate that enzyme activation by copper requires occupancy of a site in the N-terminus extension which is not present in other transport ATPases, as well as of a transmembrane site corresponding to the cation binding site of other ATPases.
摘要
ATP7B 是一种依赖铜的 P 型 ATP 酶，对铜的体内平衡至关重要。利用重组蛋白的高效异源表达，我们研究了 ATP7B 中的电荷转移。我们在添加 ATP 和形成磷酸酶中间物时检测到单个催化循环内的电荷位移。我们将这种电荷位移归因于结合在 ATP7B 内的铜的移动。基于特定的突变，我们证明铜对酶的激活需要占据 N 端延伸中的一个位点，该位点在其他转运 ATP 酶中不存在，同时还需要占据一个跨膜位点，该位点对应于其他 ATP 酶的阳离子结合位点。
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