NVT and NPT equilibration. The latter step employed the Berendsen barostat 63 as detailed above. The system was then relaxed using 5 ns of unrestrained NPT simulation using the Parrinello-Rahman barostat . Finally, a further 20 ns of NPT simulation was generated. This final 20 ns simulation was used to both derive Boresch-style 70 orientational restraint parameters and for Nwat-MM/GBSA analysis.