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ACS Synth Biol ; 8(6): 1224-1230, 2019 06 21.
Article in English | MEDLINE | ID: mdl-31051071

ABSTRACT

The assembly of channel proteins into vesicle membranes is a useful strategy to control activities of vesicle-based systems. Here, we developed a membrane AND gate that responds to both a fatty acid and a pore-forming channel protein to induce the release of encapsulated cargo. We explored how membrane composition affects the functional assembly of α-hemolysin into phospholipid vesicles as a function of oleic acid content and α-hemolysin concentration. We then showed that we could induce α-hemolysin assembly when we added oleic acid micelles to a specific composition of phospholipid vesicles. Finally, we demonstrated that our membrane AND gate could be coupled to a gene expression system. Our study provides a new method to control the temporal dynamics of vesicle permeability by controlling when the functional assembly of a channel protein into synthetic vesicles occurs. Furthermore, a membrane AND gate that utilizes membrane-associating biomolecules introduces a new way to implement Boolean logic that should complement genetic logic circuits and ultimately enhance the capabilities of artificial cellular systems.


Subject(s)
Artificial Cells , Membrane Proteins , Synthetic Biology/methods , Artificial Cells/chemistry , Artificial Cells/cytology , Artificial Cells/metabolism , Cell-Free System , Gene Regulatory Networks , Hemolysin Proteins/chemistry , Hemolysin Proteins/metabolism , Membrane Proteins/chemistry , Membrane Proteins/metabolism , Oleic Acid/chemistry , Oleic Acid/metabolism , Phosphatidylcholines/chemistry , Phosphatidylcholines/metabolism , Protein Biosynthesis
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