Biomolecular condensates sustain pH gradients at equilibrium driven by charge neutralisation

Abstract

Electrochemical gradients are essential to the functioning of cells and are typically formed across membranes using active transporters and require energy input to maintain them. Here, we show by contrast that biomolecular condensates are able to sustain significant pH gradients without any external energy input. We explore the thermodynamic driving forces that establish this gradient using a microfluidics-based droplet platform that allows us to sample in a continuous manner both the stability and composition of the condensates across a wide pH range. These results reveal that condensed biomolecular systems adjust the pH of the dense phase towards the isoelectric point (pI) of the component polypeptide chains. We demonstrate, on the basis of two representative systems, FUS and PGL3, that condensates can create both alkaline and acidic gradients with a magnitude exceeding one pH unit. Investigations of multicomponent protein/nucleic acid systems further show that heterotypic interactions can modulate condensate pH gradients. We further investigate using a bioinformatics approach the diversity of electrochemical properties of complex condensates by studying a large set of human condensate networks, showing that these span a wide range of mixture pIs and pH-response behaviours. In summary, our results reveal that protein condensation may present a fundamental physico-chemical mechanism for the effective segregation and optimisation of functional processes through changes in the emergent electrochemical microenvironment.