Engineering extrinsic disorder to control protein activity in living cells

Area of Science:

• Biochemistry
• Molecular Biology
• Cell Biology

Background:

• Optogenetics and chemogenetics enable precise control over protein signaling dynamics.
• Modulating protein structural disorder offers a novel approach to allosteric control.

Purpose of the Study:

• To develop a broadly applicable method for generating allosteric protein switches.
• To investigate the control of protein conformations and cellular morphodynamics using engineered switches.

Main Methods:

• Engineered allosteric switches by inserting light- or ligand-sensitive domains into surface-exposed protein loops.
• Computationally identified protein loops allosterically coupled to active sites.
• Introduced switches into motility signaling proteins, including kinases, guanosine triphosphatases (GTPases), and guanine exchange factors (GEFs).

Main Results:

• Successfully modulated the structural disorder of diverse proteins to create robust allosteric switches.
• Achieved controlled conversion between natural active and inactive protein conformations.
• Demonstrated modulation of living cell morphodynamics through engineered protein switches.

Conclusions:

• This approach provides a versatile strategy for designing functional protein switches.
• Engineered allosteric switches offer precise control over protein function and cellular processes.
• The method has broad applicability in biological research and synthetic biology.