To further validate our findings, we also assessed the activation energy using the PDLD/S-LRA-2000 method, as outlined in Section 2.2. The binding energies for the ground and transition states were determined by employing the substrate and a pertinent fragment of the catalytic dyad (illustrated in Scheme 1), with charges derived from charge set of the diabetic state 1. The binding energy calculations involved generating relaxed structures through MD simulations, conducted under the conditions summarized in Table . Although this study was not as extensive as the EVB study, its primary aim was to evaluate the electrostatic energies in configurations proximal to the NMR structure. Our calculations indicate a reduction in ∆∆𝑔 ‡ when transitioning from a water to an enzyme environment, yielding a value of -3.9 kcal•mol -1 (refer to Table ). This very closely replicates the EVB activation barrier and further substantiates the evidence of catalytic contribution of glucagon fibril. by averaging the corresponding ∆∆𝐺 𝑏𝑖𝑛𝑑 across 4 configurations. Each of these was taken after 20 ps of MD simulation, using 𝜀 𝑝 = 4 and charge set of the diabetic state 1. Initial configurations were produced by conducting 20 ps simulations, utilizing the reaction coordinates of the EVB surfaces of both the reactant state and the transition state.