PubAg

Main content area

Histidine Orientation Modulates the Structure and Dynamics of a de Novo Metalloenzyme Active Site

Author:
Ross, Matthew R., White, Aaron M., Yu, Fangting, King, John T., Pecoraro, Vincent L., Kubarych, Kevin J.
Source:
Journal of the American Chemical Society 2015 v.137 no.32 pp. 10164-10176
ISSN:
1520-5126
Subject:
active sites, copper, energy, histidine, hydrophobicity, infrared spectroscopy, ligands, proteins
Abstract:
The ultrafast dynamics of a de novo metalloenzyme active site is monitored using two-dimensional infrared spectroscopy. The homotrimer of parallel, coiled coil α-helices contains a His₃-Cu(I) metal site where CO is bound and serves as a vibrational probe of the hydrophobic interior of the self-assembled complex. The ultrafast spectral dynamics of Cu-CO reveals unprecedented ultrafast (2 ps) nonequilibrium structural rearrangements launched by vibrational excitation of CO. This initial rapid phase is followed by much slower ∼40 ps vibrational relaxation typical of metal-CO vibrations in natural proteins. To identify the hidden coupled coordinate, small molecule analogues and the full peptide were studied by QM and QM/MM calculations, respectively. The calculations show that variation of the histidines’ dihedral angles in coordinating Cu controls the coupling between the CO stretch and the Cu–C–O bending coordinates. Analysis of different optimized structures with significantly different electrostatic field magnitudes at the CO ligand site indicates that the origin of the stretch–bend coupling is not directly due to through-space electrostatics. Instead, the large, ∼3.6 D dipole moments of the histidine side chains effectively transduce the electrostatic environment to the local metal coordination orientation. The sensitivity of the first coordination sphere to the protein electrostatics and its role in altering the potential energy surface of the bound ligands suggests that long-range electrostatics can be leveraged to fine-tune function through enzyme design.
Agid:
5413163