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In the updated version, they improved on the energy calculations and benchmarked it against known PPIs to refine the selection criteria for an identified “hot loop” to (i) an average ΔΔ G residue ≥ 0.6 Rosetta Energy Units (REU, roughly equal to 1 kcal mol −1 ), (ii) three hot spot residues (ΔΔ G residue ≥ 0.6 REU), and (iii) a greater than 50% of binding energy contributed by the loop, represented as the sum of ΔΔ G residue
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