Microcompartmentation in Artificial Cells: pH-Induced Conformational Changes Alter Protein Localization

We report artificial cells in which protein localization in a primitive synthetic model for the cytoplasm is controlled by pH. Our model cells are giant lipid vesicles (GVs, ca. 5-30 mu m diameter) with two coexisting aqueous compartments generated by phase separation of an encapsulated poly(ethylene glycol) (PEG) and dextral) solution. Proteins arc localized to a microcompartment by partitioning between the phases. We quantified the local concentration of fluorescently labeled human serum albumin (HSA) via con focal fluorescence microscopy. At pH 6.5, the labeled HSA was more concentrated in the dextran-rich phase, but at partially/fully denaturing pH (4.1 or 12) it was localized in the PEG-rich phase. This partitioning behavior is consistent with a more expanded, hydrophobic conformation at low and high pH. Labeled VISA could be relocalized from the PEG-rich into the dextran-rich phase domain by increasing the pH from 4.1 to 6.5 to renature the protein. This approach to controlling protein localization does not require extensive reorganization of the vesicle interior: coexisting PEG-rich and dextran-rich compartments are maintained throughout the experiments. It is also quite general, we demonstrated that several other proteins varying in size and isoelectric point also relocalized within compartmentalized artificial cells in response to external pH change. This work presents stimulus-responsive protein relocalization between compartments in an artificial cell; such experimental models can provide a framework for investigating the consequences of protein localization in cell biology.