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Related Concept Videos

DNA Microarrays02:34

DNA Microarrays

Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
Proteomics01:33

Proteomics

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.
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term proteomics...
Protein Networks02:26

Protein Networks

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,...
Protein-protein Interfaces02:04

Protein-protein Interfaces

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 polypeptide...

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Related Experiment Video

Updated: Jun 14, 2026

Probing High-density Functional Protein Microarrays to Detect Protein-protein Interactions
08:07

Probing High-density Functional Protein Microarrays to Detect Protein-protein Interactions

Published on: August 2, 2015

Quantifying protein-protein interactions in high throughput using protein domain microarrays.

Alexis Kaushansky1, John E Allen, Andrew Gordus

  • 1Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.

Nature Protocols
|April 3, 2010
PubMed
Summary

Protein domain microarrays enable high-throughput identification and quantification of protein-ligand interactions. This method overcomes challenges in recombinant protein production by focusing on interaction domains, offering insights into protein-protein interaction networks.

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Identifying Protein-protein Interaction Sites Using Peptide Arrays
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Identifying Protein-protein Interaction Sites Using Peptide Arrays

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Related Experiment Videos

Last Updated: Jun 14, 2026

Probing High-density Functional Protein Microarrays to Detect Protein-protein Interactions
08:07

Probing High-density Functional Protein Microarrays to Detect Protein-protein Interactions

Published on: August 2, 2015

Extracellular Protein Microarray Technology for High Throughput Detection of Low Affinity Receptor-Ligand Interactions
06:01

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Published on: January 7, 2019

Identifying Protein-protein Interaction Sites Using Peptide Arrays
07:44

Identifying Protein-protein Interaction Sites Using Peptide Arrays

Published on: November 18, 2014

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Proteomics

Background:

  • Protein microarrays are effective for high-throughput protein-protein interaction studies.
  • Recombinant protein production can be challenging due to diverse physicochemical properties.
  • Protein interaction domains offer a more manageable approach.

Purpose of the Study:

  • To develop protocols for constructing and utilizing protein domain microarrays.
  • To quantify domain-peptide interactions efficiently.
  • To assess binding selectivity across entire domain families.

Main Methods:

  • Constructed microarrays of human Src homology 2 (SH2), phosphotyrosine binding (PTB), and mouse PDZ domains.
  • Produced domains recombinantly in Escherichia coli.
  • Quantified domain-peptide interactions using fluorescently labeled synthetic peptides and measured binding affinities (K(D)s) via saturation binding curves or fluorescence polarization.

Main Results:

  • Successfully created domain microarrays for SH2, PTB, and PDZ domains.
  • Enabled direct measurement of equilibrium dissociation constants (K(D)s) for high-affinity interactions.
  • Facilitated identification and quantification of weaker interactions using fluorescence polarization.

Conclusions:

  • Protein domain microarrays allow rapid, high-throughput identification and quantification of protein-ligand interactions with minimal sample usage.
  • Interrogating entire domain families simultaneously provides a powerful tool for assessing binding selectivity.
  • Offers an unbiased perspective on protein-protein interaction network connectivity.