Visualization of Protein Interactions in Living Cells Using Bimolecular Luminescence Complementation (BiLC)

Area of Science:

• Molecular Biology
• Biochemistry
• Cell Biology

Background:

• Intracellular protein-protein interactions (PPIs) are crucial for cellular functions and homeostasis.
• Dysregulated PPIs are implicated in various diseases and therapeutic resistance, making them potential drug targets.
• Monitoring dynamic PPIs is essential for understanding cellular processes and disease mechanisms.

Purpose of the Study:

• To describe the selection and development of bimolecular luminescence complementation (BiLC) assays.
• To demonstrate the application of BiLC assays for reporting intracellular PPIs.
• To showcase the use of BiLC in identifying small molecules that modulate PPIs.

Main Methods:

• Utilized bimolecular complementation technology, splitting a molecular reporter into two fragments.
• Fused reporter fragments to interacting proteins, reconstituting reporter activity upon PPI.
• Developed and applied bimolecular luminescence complementation (BiLC) assays to study intracellular PPIs.

Main Results:

• Successfully established BiLC assays for reporting intracellular PPIs.
• Demonstrated the utility of BiLC in identifying small molecules capable of modulating PPIs.
• Provided examples of BiLC application in drug discovery efforts.

Conclusions:

• BiLC is an effective technology for monitoring intracellular PPIs.
• BiLC assays can be employed to discover modulators of PPIs.
• This approach holds promise for identifying novel therapeutic targets and drugs.