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Protein-ligand binding kinetics are primarily controlled by the protein, not the ligand.

Srinivasan, B.; Corrionero, A.; Barone, M.; Alfonso, P.; Prendiville, N.; Cazorla, T.; Rahiyanath, A.; Whitty, A.; Tonge, P. J.

2026-05-22 biochemistry
10.64898/2026.05.20.726507 bioRxiv
Show abstract

Protein-ligand interactions underpin biological regulation and drug action, with both binding affinity and binding kinetics shaping functional outcomes. By analysing kinetic data for 4,311 protein-small-molecule pairs, we find that when association occurs below the diffusion-controlled limit, the rates of ligand association (kon) and dissociation (koff) are primarily determined by how the initial encounter complex reorganizes into the final bound state, and that this reorganization is governed chiefly by the intrinsic dynamic properties of the protein rather than by structural features of the ligand. Counterintuitively, therefore, koff exhibits minimal dependence on ligand structure, so that dissociation proceeds through protein-gated conformational transitions rather than through direct rupture of protein-ligand contacts. This mechanistic behaviour stands in marked contrast to that for protein-protein complexes, based on an analysis of 1,561 interactions. Together, these findings challenge prevailing assumptions regarding the molecular determinants of small-molecule binding kinetics, and have broad implications for rationally modulating protein-ligand interactions and drug-target residence times.

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