Protein folding is generally regarded as being driven by “hydrophobic collapse” – as the ribosome releases the nascent protein, secondary structure is rapidly acquired while tertiary structure is derived through the formation of van der Waals interactions between hydrophobic side chains and the simultaneous liberation of water molecules. Yet, in the final analysis, salt links and internal hydrogen bonds contribute nearly as much to the free energy of folding as hydrophobic forces. A key player in these hydrogen bonds is water and buried water molecules in hydrophobic interiors have the possibility of contributing to fold stability while also facilitating allostery and functional dynamics. Here, we investigate the role of hydrogen bonded water networks in allostery and function, using a homodimeric enzyme, fluoroacetate dehalogenase, as a case study. A key control in these studies involves the replacement of H2O with D2O since this has the effect of enhancing water-mediated hydrogen bond strength – an effect that is amplified exponentially in water networks. High-resolution X-ray crystallography provides an atomistic perspective of buried water networks, while subsequent Molecular Dynamics (MD) simulations and computational studies allow us to establish the hydrogen bond interaction network and the resulting allosteric contributions. 19F NMR is then used to assess fold stability, conformational transitions along the reaction coordinate, and catalytic rates. As discussed below, the results show an enhancement in allostery, cooperative dynamics, and catalysis as the water networks are reinforced by D2O. These results are discussed in the context of the potential role of buried hydrogen bonded water networks in dynamically modulating allosteric networks and response in proteins.