Moreover, the sequence between Pro28 and Pro34, which separates the α- Bias-exchange metadynamics. To complement the insights obtained with PTMetaD-WTE, we adopted BEMD method to further explore the conformational ensemble of DHH1N. We constructed 1D-FE profiles for CHARMM36m and CHARMM22* at 300 K and compare them with the results from PTMetaD-WTE. Similarly, the configurational ensemble projected in most CVs also displays a single FE minimum (Fig. To further examine the conformational states of DHH1N, we construct 2D-FES for CHARMM36m and CHARMM22* BEMD and compare them with the 2D-FES from PTMetaD-WTE. Similarly, every simulation reveals a global minimum at low αcontent in the disordered cluster 1, accompanied by multiple shallow local minima, indicating a large number of substates as a result of conformational heterogeneity (Fig. ). Cluster 1 of CHARMM36m and CHARMM22* BEMD comprises approximately 70 % of the equilibrium conformational population, similar to the result of CHARMM36m PTMetaD-WTE, while cluster 1 population of CHARMM22* PTMetaD-WTE is much lower due to relatively shorter simulation time (Supplementary Note 5, Table ). The largest disparities between the 2D-FES are primarily due to different choices of force-fields. In detail, the CHARMM36m results