Tanchou Valérie, Gas Fabienne, Urvoas Agathe, Cougouluègne Françoise, Ruat Sylvie, Averseng Olivier, Quéméneur Eric
CEA VALRHO, Département d'Ingénierie et d'Etudes des Protéines, Service de Biochimie Post-génomique et Toxicologie Nucléaire, BP1717, 30207 Bagnols sur Cèze cedex, France.
Biochem Biophys Res Commun. 2004 Dec 10;325(2):388-94. doi: 10.1016/j.bbrc.2004.10.048.
The HAH1 metallochaperone is a key protein implicated in copper homeostasis in human cells. Using as solid-phase based assay completed with Biacore studies, we provided evidence that HAH1 forms homo-dimers in the presence of copper. Biacore analysis allowed us to determine the kinetic parameters of this interaction, characterised by an apparent affinity constant of 6muM. Moreover, we demonstrated that copper-loaded HAH1 interacts independently with each of the six individual metal-binding domains of the copper-translocating Menkes ATPase. Finally, the homo-dimerisation of the metallochaperone was confirmed in living cells by using fluorescence resonance energy transfer. Results have been discussed in the context of intracellular copper control.
HAH1金属伴侣蛋白是一种在人类细胞铜稳态中起关键作用的蛋白质。通过使用基于固相分析并结合Biacore研究，我们提供了证据表明HAH1在铜存在的情况下形成同源二聚体。Biacore分析使我们能够确定这种相互作用的动力学参数，其特征在于表观亲和常数为6μM。此外，我们证明了负载铜的HAH1与铜转运门克斯ATP酶的六个单独的金属结合结构域中的每一个独立相互作用。最后，通过使用荧光共振能量转移在活细胞中证实了金属伴侣蛋白的同源二聚化。已在细胞内铜控制的背景下讨论了结果。
