How conformational changes can affect catalysis, inhibition and drug resistance of enzymes with induced-fit binding mechanism such as the HIV-1 protease

TL;DR

This paper presents a general model for how conformational changes in enzymes with induced-fit mechanisms, like HIV-1 protease, influence catalysis, inhibition, and drug resistance, especially considering mutations that shift conformational equilibria.

Contribution

It introduces a mathematical framework linking conformational dynamics to enzyme activity and drug resistance, highlighting the impact of mutations on these processes.

Findings

Mutations destabilizing the closed conformation increase catalytic rates in the presence of inhibitors.

The effect of conformational shifts on catalysis is independent of the inhibitor molecule.

Experimental data for HIV-1 protease mutation L90M supports the model's predictions.

Abstract

A central question is how the conformational changes of proteins affect their function and the inhibition of this function by drug molecules. Many enzymes change from an open to a closed conformation upon binding of substrate or inhibitor molecules. These conformational changes have been suggested to follow an induced-fit mechanism in which the molecules first bind in the open conformation in those cases where binding in the closed conformation appears to be sterically obstructed such as for the HIV-1 protease. In this article, we present a general model for the catalysis and inhibition of enzymes with induced-fit binding mechanism. We derive general expressions that specify how the overall catalytic rate of the enzymes depends on the rates for binding, for the conformational changes, and for the chemical reaction. Based on these expressions, we analyze the effect of mutations that mainly shift the conformational equilibrium on catalysis and inhibition. If the overall catalytic rate is limited by product unbinding, we find that mutations that destabilize the closed conformation relative to the open conformation increase the catalytic rate in the presence of inhibitors by a factor exp(ddG/RT) where ddG is the mutation-induced shift of the free-energy difference between the conformations. This increase in the catalytic rate due to changes in the conformational equilibrium is independent of the inhibitor molecule and, thus, may help to understand how non-active-site mutations can contribute to the multi-drug-resistance that has been observed for the HIV-1 protease. A comparison to experimental data for the non-active-site mutation L90M of the HIV-1 protease indicates that the mutation slightly destabilizes the closed conformation of the enzyme.