Probing the Coordination Environment of the Human Copper Chaperone HAH1: Characterization of HgII-Bridged Homodimeric Species in Solution

• Published in: Chemistry : a European journal Vol. 19; no. 27; pp. 9042 - 9049
• Main Authors: Łuczkowski, Marek, Zeider, Brian A., Hinz, Alia V. H., Stachura, Monika, Chakraborty, Saumen, Hemmingsen, Lars, Huffman, David L., Pecoraro, Vincent L.
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 01.07.2013; WILEY‐VCH Verlag; Wiley Subscription Services, Inc
• Subjects: 199mHg PAC; Chemistry; copper; mercury; metallochaperones; NMR spectroscopy
• ISSN: 0947-6539; 1521-3765
• DOI: 10.1002/chem.201204184

Although metal ion homeostasis in cells is often mediated through metallochaperones, there are opportunities for toxic metals to be sequestered through the existing transport apparatus. Proper trafficking of CuI in human cells is partially achieved through complexation by HAH1, the human metallochaperone responsible for copper delivery to the Wilson and Menkes ATPase located in the trans‐Golgi apparatus. In addition to binding copper, HAH1 strongly complexes HgII, with the X‐ray structure of this complex previously described. It is important to clarify the solution behavior of these systems and, therefore, the binding of HgII to HAH1 was probed over the pH range 7.5 to 9.4 using 199Hg NMR, 199mHg PAC and UV–visible spectroscopies. The metal‐dependent protein association over this pH range was examined using analytical gel‐filtration. It can be concluded that at pH 7.5, HgII is bound to a monomeric HAH1 as a two coordinate, linear complex (HgS2), like the HgII–Atx1 X‐ray structure (PDB ID: 1CC8). At pH 9.4, HgII promotes HAH1 association, leading to formation of HgS3 and HgS4 complexes, which are in exchange on the μs–ns time scale. Thus, structures that may represent central intermediates in the process of metal ion transfer, as well as their exchange kinetics have been characterized. HAH1, the HgII carrier? Trafficking of CuI in human cells is achieved through complexation by the metallochaperone, HAH1. In addition, HAH1 strongly binds HgII; the solution behavior of HgII–HAH1 complexation (see figure) has remained obscure, but is elucidated in this work. Structures are reported that may represent central intermediates in the processing of HgII ions.