Advances in proximity labeling strategies for interactome mapping and functional interrogation

Area of Science:

• Molecular Biology
• Biochemistry
• Cell Biology

Background:

• Classical biochemical methods struggle to capture transient or weak protein-protein interactions (PPIs) under native conditions.
• Proximity labeling (PL) overcomes these limitations by enabling in situ covalent tagging of proximal molecules within intact biological systems.
• PL provides spatiotemporally resolved insights into cellular molecular organization.

Purpose of the Study:

• To review recent significant methodological and conceptual advances in proximity labeling technologies.
• To highlight how these innovations expand the scope of interactome analysis.
• To discuss future opportunities and challenges in the field.

Main Methods:

• Review of recent literature on enzymatic, chemical, and photocatalytic proximity labeling platforms.
• Discussion of hybrid genetic-chemical and optogenetic strategies for PL.
• Focus on advances enabling microenvironment-specific targeting and reduced background.

Main Results:

• Recent PL advancements offer improved control over labeling radius, kinetics, and activation.
• New PL strategies facilitate the discovery of previously inaccessible interaction networks, including membrane assemblies and chromatin complexes.
• PL enables the study of in vivo protein microenvironments.

Conclusions:

• Proximity labeling has revolutionized interactome mapping by providing native snapshots of molecular neighborhoods.
• Continued innovation in PL platforms promises deeper understanding of cellular organization and function.
• Future directions include refining PL for greater precision and exploring its potential in signal amplification.