Single-molecule theory of enzymatic inhibition predicts the emergence of inhibitor-activator duality

TL;DR

This paper revises classical enzymatic inhibition theory from a single-molecule perspective, revealing conditions where inhibitors can paradoxically activate enzymes, especially in uncompetitive and mixed inhibition modes, due to stochastic enzyme fluctuations.

Contribution

It introduces a new stochastic framework for enzyme inhibition, predicting inhibitor-activator duality and providing measurable conditions for its emergence.

Findings

Classical results hold for competitive inhibition despite enzyme complexity.

Stochastic fluctuations can cause inhibitors to facilitate enzyme activity in certain modes.

Mathematical conditions for inhibitor-activator duality are derived and experimentally relevant.

Abstract

The classical theory of enzymatic inhibition aims to quantitatively describe the effect of certain molecules -- called inhibitors -- on the progression of enzymatic reactions, but growing signs indicate that it must be revised to keep pace with the single-molecule revolution that is sweeping through the sciences. Here, we take the single enzyme perspective and rebuild the theory of enzymatic inhibition from the bottom up. We find that accounting for multi-conformational enzyme structure and intrinsic randomness cannot undermine the validity of classical results in the case of competitive inhibition; but that it should strongly change our view on the uncompetitive and mixed modes of inhibition. There, stochastic fluctuations on the single-enzyme level could give rise to inhibitor-activator duality -- a phenomenon in which, under some conditions, the introduction of a molecule whose binding shuts down enzymatic catalysis will counter intuitively work to facilitate product formation. We state -- in terms of experimentally measurable quantities -- a mathematical condition for the emergence of inhibitor-activator duality, and propose that it could explain why certain molecules that act as inhibitors when substrate concentrations are high elicit a non-monotonic dose response when substrate concentrations are low. The fundamental and practical implications of our findings are thoroughly discussed.