Proximity-dependent labeling methods for proteomic profiling in living cells: An update.

Area of Science:

• Cellular Biology
• Proteomics
• Biochemistry

Background:

• Understanding cellular organization and protein interactions is crucial for deciphering biological processes.
• Spatially restricted proteome analysis requires advanced techniques to map protein localization and interactions within living cells.

Purpose of the Study:

• To provide an updated description and comparison of proximity labeling methods.
• To highlight the applications and improvements of proximity labeling technologies.
• To guide researchers in selecting appropriate proximity labeling methods for specific biological questions.

Main Methods:

• Proximity labeling utilizes enzymes to generate reactive radicals, covalently tagging neighboring proteins within living cells.
• Mass spectrometry (MS) is employed to isolate and analyze the tagged endogenous proteins.
• Enzyme fusion with specific proteins or signal peptides enables spatial targeting to particular subcellular regions.

Main Results:

• Proximity labeling coupled with MS offers a high-throughput approach for systematic analysis of spatially restricted proteomes.
• These technologies have provided significant insights into diverse biological processes.
• Different proximity labeling methods possess unique features suitable for addressing distinct biological inquiries.

Conclusions:

• Proximity labeling is a powerful tool for investigating cellular organization and protein interactome networks.
• The choice of proximity labeling method depends on the specific biological question being addressed.
• Continued advancements in proximity labeling promise deeper understanding of cellular functions.