Cu(I) Controls Conformational States in Human Atox1 Metallochaperone: An EPR and Multiscale Simulation Study

• DOI: 10.1021/acs.jpcb.0c01744
• Publication Date: 2020-05-12T20:53:05Z
• Authors: Ortal Perkal; Zena Qasem; Meital Turgeman; Renana Schwartz; Lada Gevorkyan-Ai...; Matic Pavlin; Alessandra Magist...; Dan Thomas Major; Sharon Ruthstein
• Keywords: Metallochaperones; Binding Sites; Copper Transport Proteins; Humans; molecular dynamics, Copper, EPR, Atox1; 01 natural sciences; Copper; Molecular Chaperones; 0104 chemical sciences

Atox1 is a human copper metallochaperone that is responsible for transferring copper ions from the main human copper transporter, hCtr1, to ATP7A/B in the Golgi apparatus. Atox1 interacts with the Ctr1 C-terminal domain as a dimer, although it transfers the copper ions to ATP7A/B in a monomeric form. The copper binding site in the Atox1 dimer involves Cys12 and Cys15, while Lys60 was also suggested to play a role in the copper binding. We recently showed that Atox1 can adopt various conformational states, depending on the interacting protein. In the current study, we apply EPR experiments together with hybrid quantum mechanics-molecular mechanics molecular dynamics simulations using a recently developed semiempirical density functional theory approach, to better understand the effect of Atox1's conformational states on copper coordination. We propose that the flexibility of Atox1 occurs owing to protonation of one or more of the cysteine residues, and that Cys15 is an important residue for Atox1 dimerization, while Cys12 is a critical residue for Cu(I) binding. We also show that Lys60 electrostatically stabilizes the Cu(I)-Atox1 dimer.