Proton network flexibility enables robustness and large electric fields in the ketosteroid isomerase active site

TL;DR

This study combines advanced simulations to reveal how hydrogen bond networks in ketosteroid isomerase's active site are robust and generate large electric fields, crucial for enzyme function.

Contribution

It demonstrates the proton and electronic flexibility of hydrogen bond networks in enzyme active sites using ab initio simulations, explaining their robustness and electric field effects.

Findings

Hydrogen bond network responds to mutations by shifting protons and redistributing charge.

Proton flexibility increases with extended hydrogen bond networks and analog presence.

Large electric fields in the active site are explained by network dynamics and charge redistribution.

Abstract

Hydrogen bond networks play vital roles in biological functions ranging from protein folding to enzyme catalysis. Here we combine electronic structure calculations and ab initio path integral molecular dynamics simulations, which incorporate both nuclear and electronic quantum effects, to show why the network of short hydrogen bonds in the active site of ketosteroid isomerase is remarkably robust to mutations along the network and how this gives rise to large local electric fields. We demonstrate that these properties arise from the network's ability to respond to a perturbation by shifting proton positions and redistributing electronic charge density. This flexibility leads to small changes in properties such as the partial ionization of residues and $pK_a$ isotope effects upon mutation of the residues, consistent with recent experiments. This proton flexibility is further enhanced when an extended hydrogen bond network forms in the presence of an intermediate analog, which allows us to explain the chemical origins of the large electric fields in the enzyme's active site observed in recent experiments.