Reversibly Triggered Protein-Ligand Assemblies in Giant Vesicles

Area of Science:

• Biochemistry
• Synthetic Biology
• Biophysics

Background:

• Dynamic control over molecular interactions is crucial for developing advanced biomimetic systems.
• Giant vesicles serve as model systems for cellular compartments, enabling the study of complex biological processes.

Purpose of the Study:

• To establish a method for externally controlling protein-ligand interactions within giant vesicles.
• To demonstrate reversible modulation of these interactions using small-molecule triggers.

Main Methods:

• Utilized alcohol dehydrogenase to alter the internal pH of giant vesicles.
• Employed pH-sensitive interactions between a nickel-nitrilotriacetic acid (Ni-NTA) ligand and an oligohistidine-tagged protein.
• Alternated the addition of different small-molecule substrates to control pH and protein-ligand binding.

Main Results:

• Successfully demonstrated reversible control over protein-ligand interactions in giant vesicles.
• Showcased the ability to modulate these interactions by adjusting the internal vesicle pH via enzymatic conversion of small molecules.
• Established a system for dynamic and externally triggered molecular assembly and disassembly.

Conclusions:

• External small-molecule triggers can effectively and reversibly control dynamic protein-ligand interactions in giant vesicles.
• This pH-modulation approach offers a versatile tool for synthetic biology and the construction of responsive biomimetic systems.
• The findings pave the way for designing more sophisticated artificial cells and dynamic molecular devices.