Design of Light-Controlled Protein Conformations and Functions

Area of Science:

• Biochemistry and Molecular Biology
• Chemical Biology
• Protein Engineering

Background:

• Controlling protein function with light offers unique advantages like spatial and temporal precision.
• Existing methods for protein control lack the selectivity and fine-tuning capabilities of light-based approaches.
• Interest in optogenetic tools for manipulating biological systems is rapidly growing.

Purpose of the Study:

• To describe a generalizable protocol for engineering proteins to be light-controllable.
• To provide a framework for selecting suitable proteins and modification sites for light-induced control.
• To demonstrate the protocol's utility with examples from existing literature.

Main Methods:

• Development of a protocol for protein engineering using exogenously introduced photoisomerizable small molecules.
• Application of the protocol to modify the calcium affinity of cadherin, a cell-cell adhesion protein.
• Application of the protocol to engineer the opening and closing of the chaperonin Mm-cpn.

Main Results:

• Successful engineering of cadherin to switch between native and disrupted states using light-induced small molecule reactions.
• Successful engineering of Mm-cpn to switch between functional states (open/closed) via light-activated small molecules.
• Demonstration of a systematic approach for protein light-control engineering, from protein selection to functional assays.

Conclusions:

• The described protocol provides a versatile method for achieving light-based control over diverse protein functions.
• This approach enables precise manipulation of protein activity, advancing the field of chemical biology and optogenetics.
• The protocol serves as a guide for researchers aiming to develop novel light-switchable protein systems.