Dynamic Allostery of the Catabolite Activator Protein Revealed by Interatomic Forces

Maxime Louet, Christian Seifert, Ulf Hensen, Frauke Gräter

Abstract: Author Summary The Catabolite Activator Protein (CAP) is a well-studied example for how cellular catabolite levels are integrated into the gene regulation. Its affinity for a specific stretch of DNA can be switched on by the binding of two nucleotide molecules termed cAMP to its two protomers. Even though the nucleotides occupy structurally identical binding pockets, the second cAMP binding occurs at an affinity orders of magnitude lower than the first cAMP binding. The question arises how, in the absence of structural changes, the first binding can affect the second. An answer from experiments has been that the communication is largely of entropic nature, i.e. the second cAMP binding would lead to a pronounced reduction in atomic fluctuations of the protein without affecting the atomic mean positions. We here revisited this question by performing Molecular Dynamics simulations. By measuring correlations of forces, a newly derived method outperforming the more common coordinate-based approach, we could recover the previously determined entropic penalty. In addition, however, we observed unobtrusive structural changes of side-chain interactions leading to the occlusion of the second binding pocket that add a critical ‘enthalpic’ component hitherto overlooked. Our study provides a mechanistic view onto the intriguing anti-cooperativity of CAP.