Riboswitch-controlled lipid conversion enables functional membrane asymmetry in artificial cells

Area of Science:

• Synthetic biology
• Biochemistry
• Materials science

Background:

• Dynamic lipid membrane regulation is crucial for cellular functions.
• Synthetic analogs for controlled membrane remodeling are limited.
• Artificial cells require stimulus-responsive components.

Purpose of the Study:

• To develop an artificial cell platform for stimulus-responsive lipid membrane remodeling.
• To enable dynamic changes in membrane composition via gene expression.
• To create functional artificial cells with programmable properties.

Main Methods:

• Utilized a fluoride-responsive riboswitch to control phospholipase D (PLD) expression.
• Employed cell-free synthesis of PLD within lipid vesicles.
• Hydrolyzed phosphatidylcholine (PC) to phosphatidic acid (PA) to induce membrane asymmetry.
• Functionalized membranes with mechanosensitive channels (e.g., MscL).

Main Results:

• Demonstrated riboswitch-mediated, stimulus-responsive lipid remodeling in artificial cells.
• Generated negatively charged membrane asymmetry through enzymatic hydrolysis.
• Successfully incorporated mechanosensitive channels into remodeled membranes.
• Characterized the kinetics of asymmetry generation under varying conditions.

Conclusions:

• The platform couples gene expression to dynamic membrane remodeling.
• Riboswitch-regulated lipid transitions are key for environment-responsive artificial cells.
• This approach offers a strategy for building artificial cells with programmable functions.