In summary, while the same f opt 3 can be achieved through different combinations of K A-L , K B-L , and α (e.g., contour lines in Figure and), higher cooperativity α with weaker binding affinities can reach an equally high f opt 3 with a milder hook effect (Figure and Figure in the Supporting Information), making cooperativity an attractive optimization target even at the cost of a weakened binding affinity.