Spectroscopic and electronic structure studies of the trinuclear Cu cluster active site of the multicopper oxidase laccase: nature of its coordination unsaturation.

Abstract

Laccase is a multicopper oxidase that contains four Cu ions, one type 1 (T1), one type 2 (T2), and a coupled binuclear type 3 Cu pair (T3). The T2 and T3 centers form a trinuclear Cu cluster that is the active site for O2 reduction to H2O. A combination of spectroscopic and DFT studies on a derivative where the T1 Cu has been replaced by a spectroscopically innocent Hg2+ ion has led to a detailed geometric and electronic structure description of the resting trinuclear Cu cluster, complementing crystallographic results. The nature of the T2 Cu ligation has been elucidated; this site is three-coordinate with two histidines and a hydroxide over its functional pH range (stabilized by a large inductive effect, cluster charge, and a hydrogen-bonding network). Both the T2 and T3 Cu centers have open coordination positions oriented toward the center of the cluster. DFT calculations show that the negative protein pocket (four conserved Asp/Glu residues within 12 A) and the dielectric of the protein play important roles in the electrostatic stability and integrity of the highly charged, coordinatively unsaturated trinuclear cupric cluster. These tune the ligand binding properties of the cluster, leading to its high affinity for fluoride and its coordination unsaturation in aqueous media, which play a key role in its O2 reactivity.

Cite this paper

@article{Quintanar2005SpectroscopicAE, title={Spectroscopic and electronic structure studies of the trinuclear Cu cluster active site of the multicopper oxidase laccase: nature of its coordination unsaturation.}, author={Liliana Quintanar and Jungjoo Yoon and Constantino P Aznar and Amy E Palmer and Kristoffer K. Andersson and R David Britt and Edward I. Solomon}, journal={Journal of the American Chemical Society}, year={2005}, volume={127 40}, pages={13832-45} }