Benchmarking biomolecular force field-based Zn²⁺ for mono- and bimetallic ligand binding sites

Zn²⁺ is one of the most versatile biologically available metal ions, but accurate modeling of Zn²⁺-containing metalloproteins at the biomolecular force field level can be challenging. Since most Zn²⁺ models are parameterized in bulk solvent, in-depth knowledge about their performance in a protein environment is limited. Thus, we systematically investigate here the behavior of non-polarizable Zn²⁺ models for their ability to reproduce experimentally determined metal coordination and ligand binding in metalloproteins. The benchmarking is performed in challenging environments, including mono- (carbonic anhydrase II) and bimetallic (metallo-β-lactamase VIM-2) ligand binding sites. We identify key differences in the performance between the Zn²⁺ models with regard to the preferred ligating atoms (charged/non-charged), attraction of water molecules, and the preferred coordination geometry. Based on these results, we suggest suitable simulation conditions for varying Zn²⁺ site geometries that could guide the further development of biomolecular Zn²⁺ models.