Techniques for detecting protein-protein interactions in living cells: principles, limitations, and recent progress

Area of Science:

• Biochemistry
• Cell Biology
• Molecular Biology

Background:

• Protein-protein interactions (PPIs) are vital for cellular processes like signal transduction.
• Traditional biochemical methods lack the spatio-temporal resolution needed for in vivo studies.
• Detecting PPIs in living cells requires advanced techniques.

Purpose of the Study:

• To review and summarize key in vivo techniques for detecting protein-protein interactions.
• To focus on the principles, applications, advantages, and limitations of these methods.
• To introduce novel techniques and recent advancements in PPI detection.

Main Methods:

• Review of established and emerging in vivo techniques for PPI detection.
• Focus on Fluorescence Resonance Energy Transfer (FRET)-based methods, including single-molecule FRET (smFRET) and FRET-fluorescence lifetime imaging microscopy (FRET-FLIM).
• Discussion of cytoskeleton-based assay for protein-protein interaction (CAPPI) and single-molecule protein proximity index (smPPI).

Main Results:

• Summary of three main categories of in vivo PPI detection techniques.
• Detailed comparison of the advantages and limitations of each method.
• Highlighting of recent improvements and novel approaches in the field.

Conclusions:

• The review provides a comprehensive reference for selecting appropriate in vivo PPI detection techniques.
• Understanding the nuances of each method is crucial for successful biological studies.
• Advancements in techniques offer greater insight into molecular interactions within living cells.