Simulations of a protein fold switch reveal crowding-induced population shifts driven by disordered regions

Abstract

Macromolecular crowding effects on globular proteins, which usually adopt a single stable fold, have been widely studied. However, little is known about crowding effects on fold-switching proteins, which reversibly switch between distinct folds. Here we study the mutationally driven switch between the folds of GA and GB, the two 56-amino acid binding domains of protein G, using a structure-based dual-basin model. We show that, in the absence of crowders, the fold populations PA and PB can be controlled by the strengths of contacts in the two folds, κA and κB. A population balance, PA ≈ PB, is obtained for κB/κA = 0.92. The resulting model protein is subject to crowding at different packing fractions, ϕc. We find that crowding increases the GB population and reduces the GA population, reaching PB/PA ≈ 4 at ϕc = 0.44. We analyze the ϕc-dependence of the crowding-induced GA-to-GB switch using scaled particle theory, which provides a qualitative, but not quantitative, fit of our data, suggesting effects beyond a spherical description of the folds. We show that the terminal regions of the protein chain, which are intrinsically disordered only in GA, play a dominant role in the response of the fold switch to crowding effects.