Kinetic network model to explain gain-of-function mutations in ERK2 enzymes

Abstract

ERK2 is a kinase protein that belongs to a Ras/Raf/MEK/ERK signalling pathway, which is activated in response to a range of extracellular signals. Malfunctioning of this cascade leads to variety of serious diseases, including cancers. This is often caused by mutations in proteins belonging to the cascade, frequently leading to abnormally high activity of the cascade even in the absence of external signal. One such gain-of-function mutation in ERK2 protein, called a sevenmaker mutation (D319N), was discovered in 1994 in Drosophila. This mutation leads to disruption of interactions of other proteins with D-site of ERK2 and results, contrary to expectations, in increase of its activity in vivo. However, no molecular mechanism to explain this effect has been presented so far. The difficulty is that this mutation should equally negatively affect interactions of ERK2 with all substrates, activators and deactivators. In this paper, we present a quantitative kinetic network model that gives a possible explanation of the increased activity of mutant ERK2 species. A simplified biochemical network for ERK2, viewed as a system of coupled Michaelis-Menten processes, is presented. Its dynamic properties are calculated explicitly using the method of first-passage processes. The effect of mutation is associated with changes in the strength of interaction energy between the enzyme and the substrates. It is found that the dependence of kinetic properties of the protein on the interaction energy is non-monotonic, suggesting that some mutations might lead to more efficient catalytic properties, despite weakening inter-molecular interactions. Our theoretical predictions agree with experimental observations for the sevenmaker mutation in ERK2. It is also argued that the effect of mutations might depend on the concentrations of substrates.