Mechanisms of Copper Selectivity and Release by the Metallochaperone CusF: Insights from CO-Binding, Rapid-Freeze-Quench EXAFS, and Unnatural Amino Acid Substitution.

ABSTRACT: Metallochaperones are small proteins that shuttle essential metal ions such as Cu selectively to their cellular targets. CusF has unusual Cu(I) coordination, bound by two methionines, one histidine and a capping tryptophan residue, W44. Here we compare the CO binding reactivity of the wild type (WT) protein and its W44A, F, and M variants. Fourier-transform infrared (FTIR) indicates that W44A provides unhindered access for CO, while W44M is unreactive. WT is also largely unreactive to CO suggesting that the tryptophan cap is effective in shielding the Cu(I) center from exogenous adduct formation, while the Phe variant shows partial reactivity suggestive of an equilibrium between cap-on and cap-off conformers. Rates of metal transfer to the partner CusB are consistent with the π-cation cap

SUBMITTER: Rush KW 

PROVIDER: S-EPMC11611273 | biostudies-literature | 2024 Nov

REPOSITORIES: biostudies-literature