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Related Concept Videos

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Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
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DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
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Related Experiment Video

Updated: Dec 9, 2025

Characterization of Neuronal Lysosome Interactome with Proximity Labeling Proteomics
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Proximity-dependent labeling methods for proteomic profiling in living cells: An update.

Justin A Bosch1, Chiao-Lin Chen1, Norbert Perrimon1,2

  • 1Department of Genetics, Blavatnik Institute, Harvard Medical School, Boston, Massachusetts, USA.

Wiley Interdisciplinary Reviews. Developmental Biology
|September 10, 2020
PubMed
Summary

Proximity labeling techniques coupled with mass spectrometry enable high-throughput analysis of cellular proteomes. This review compares methods for understanding protein interactions and subcellular localization in living cells.

Keywords:
APEXBioIDHRPproximity labeling PUP-IT

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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.