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Related Concept Videos

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

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Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays
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Published on: October 23, 2019

Using the Nucleic Acid Programmable Protein Array (NAPPA) for Identifying Protein-Protein Interactions: General

Andrew J Link1, Joshua Labaer

  • 1Vanderbilt University School of Medicine, Nashville, TN 37232-8575, USA.

CSH Protocols
|March 2, 2011
PubMed
Summary

Nucleic Acid Programmable Protein Array (NAPPA) enables protein microarray production. This study explores two interaction detection schemas, highlighting the benefits of coexpressing a query protein on the NAPPA slide.

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Last Updated: Jun 4, 2026

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08:07

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

Published on: August 2, 2015

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Proteomics

Background:

  • Protein microarrays are crucial for studying protein interactions.
  • Nucleic Acid Programmable Protein Array (NAPPA) is a method for in situ protein production on microarrays.
  • Identifying protein interactions is essential for understanding biological pathways.

Purpose of the Study:

  • To describe the advantages of using a coexpressed query protein in Nucleic Acid Programmable Protein Array (NAPPA) experiments.
  • To provide guidance on selecting appropriate epitope tags for coexpressed query proteins.
  • To compare two general schemas for identifying protein interactions using NAPPA.

Main Methods:

  • Utilizing cell-free extracts for in situ transcription and translation of cDNAs on glass slides.
  • Probing expressed NAPPA slides with purified query proteins.
  • Coexpressing query proteins simultaneously with target proteins on the NAPPA slide.

Main Results:

  • The first schema involves probing with purified query proteins, allowing for varied binding conditions and post-translational modifications.
  • The second schema involves coexpressing the query protein on the NAPPA slide.
  • Both schemas allow for signal detection via direct labeling or antibody-based detection.

Conclusions:

  • Coexpression of query proteins offers distinct advantages for protein interaction studies using NAPPA.
  • Careful selection of epitope tags is important for successful coexpression strategies.
  • NAPPA technology provides a flexible platform for protein interaction discovery.