Constructing Photoactivatable Protein with Genetically Encoded Photocaged Glutamic Acid.

Area of Science:

• Biochemistry
• Molecular Biology
• Synthetic Biology

Background:

• Genetic code expansion (GCE) allows creating photoactivatable proteins by replacing residues with photocaged analogues.
• Current limitations exist due to a restricted set of photocaged amino acids available for GCE.

Purpose of the Study:

• To genetically incorporate photocaged glutamic acid (Glu) analogues into proteins.
• To demonstrate the utility of photocaged Glu in creating novel photoactivatable proteins.

Main Methods:

• Utilized genetic code expansion (GCE) in both E. coli and mammalian cells.
• Introduced photocaged glutamic acid analogues at specific protein sites.
• Constructed photoactivatable variants of fluorescent proteins and SpyCatcher.

Main Results:

• Successfully achieved genetic incorporation of photocaged Glu analogues in diverse cell types.
• Demonstrated the generation of functional photoactivatable proteins using the new GCE strategy.
• Showcased applications in creating light-switchable fluorescent proteins and SpyCatcher.

Conclusions:

• Genetically encoded photocaged Glu significantly expands the available amino acid repertoire for GCE.
• This method provides a powerful tool for designing and applying photoactivatable proteins.
• Opens new avenues for controlling protein function with light in biological systems.