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Related Concept Videos

Protein Networks02:26

Protein Networks

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An organism can have thousands of different proteins, and these proteins must cooperate to ensure the health of an organism. Proteins bind to other proteins and form complexes to carry out their functions. Many proteins interact with multiple other proteins creating a complex network of protein interactions.
These interactions can be represented through maps depicting protein-protein interaction networks, represented as nodes and edges. Nodes are circles that are representative of a protein,...
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Protein-protein Interfaces02:04

Protein-protein Interfaces

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Many proteins form complexes to carry out their functions, making protein-protein interactions (PPIs) essential for an organism's survival. Most PPIs are stabilized by numerous weak noncovalent chemical forces. The physical shape of the interfaces determines the way two proteins interact. Many globular proteins have closely-matching shapes on their surfaces, which form a large number of weak bonds. Additionally, many PPIs occur between two helices or between a surface cleft and a...
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Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
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Proteomics01:33

Proteomics

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A proteome is the entire set of proteins that a cell type produces. We can study proteomes using the knowledge of genomes because genes code for mRNAs, and the mRNAs encode proteins. Although mRNA analysis is a step in the right direction, not all mRNAs are translated into proteins.
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Related Experiment Video

Updated: Apr 26, 2026

Discovering Protein Interactions and Characterizing Protein Function Using HaloTag Technology
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Discovering Protein Interactions and Characterizing Protein Function Using HaloTag Technology

Published on: July 12, 2014

38.9K

Discovering protein interactions and characterizing protein function using HaloTag technology.

Danette L Daniels1, Jacqui Méndez2, Hélène Benink2

  • 1Promega Corporation; Danette.Daniels@promega.com.

Journal of Visualized Experiments : Jove
|July 22, 2014
PubMed
Summary
This summary is machine-generated.

HaloTag technology enables efficient isolation of human protein complexes, advancing proteomic analysis. This covalent capture method improves discovery of protein interactions and cellular localization for eukaryotes.

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Discovering Protein Interactions and Characterizing Protein Function Using HaloTag Technology
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A Protocol for the Identification of Protein-protein Interactions Based on 15N Metabolic Labeling, Immunoprecipitation, Quantitative Mass Spectrometry and Affinity Modulation
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Area of Science:

  • Proteomics
  • Molecular Biology
  • Biochemistry

Background:

  • Human proteome analysis lags due to protein number and interaction complexity.
  • Traditional methods struggle with weak, transient, or low-abundance protein interactions.

Purpose of the Study:

  • Develop HaloTag technology for efficient human protein complex isolation.
  • Characterize protein function, interactions, and cellular localization in eukaryotes.

Main Methods:

  • Genetically encoded HaloTag fusion protein for covalent immobilization.
  • Protein pull-down assays for interaction discovery and functional assays.
  • Cellular localization studies.

Main Results:

  • HaloTag offers superior speed, specificity, and covalent capture compared to non-covalent methods.
  • Successfully isolated and characterized epigenetic proteins BRD4 and HDAC1.
  • Demonstrated utility in discovering novel interactions and determining cellular localization.

Conclusions:

  • HaloTag technology significantly enhances human proteomic analysis.
  • Enables efficient discovery of novel protein interactions and characterization of cellular localization.
  • Advances understanding of human functional proteomics through improved protein isolation techniques.