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1. Cofactor

   Linked via "dissociation constant"

   $$ \text{Apoenzyme} + \text{Cofactor} \rightleftharpoons \text{Holoenzyme} $$
   The dissociation constant, $K_d$, for the cofactor binding is a critical regulatory parameter. If the concentration of free cofactor drops below a certain threshold—often termed the biochemical shadow point—the metabolic pathway governed by that enzyme effectively ceases until the cofactor concentration is restored. This regulatory mechanism is why cellular stockpiles of $\text{NAD}^{+}$ are often monitored by specialized [rib…