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Assessment of Resistance to Tyrosine Kinase Inhibitors by an Interrogation of Signal Transduction Pathways by Antibody Arrays
07:42

Assessment of Resistance to Tyrosine Kinase Inhibitors by an Interrogation of Signal Transduction Pathways by Antibody Arrays

Published on: September 19, 2018

SH2 domain-based tyrosine phosphorylation array

Xin Jiang1, Lesile Roth, Stephanie Han

  • 1Panomics, Inc., Herts, 6519 Dumbarton Cr., Freemont, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|March 29, 2008
PubMed

Insights

This study introduces a new high-throughput assay to detect protein interactions. This method profiles cellular phosphorylation status by analyzing interactions between phosphorylated proteins and Src homology-2 (SH2) domains.

Area of Science:

  • Cellular biology
  • Molecular biology
  • Biochemistry

Background:

  • Tyrosine phosphorylation is crucial for intracellular signal transduction.
  • Phosphorylated tyrosine residues recruit proteins with Src homology-2 (SH2) domains to initiate signaling pathways.

Purpose of the Study:

  • To develop a high-throughput assay for detecting interactions between phosphorylated proteins and human SH2 domains.
  • To profile the genome-wide phosphorylation status of cells under various biological conditions.

Main Methods:

  • An SH2 domain array was created by immobilizing 115 SH2 domain proteins on a membrane.
  • The assay involves incubating cell lysate with the array membrane.
  • Signal detection uses a chemiluminescence-based system, requiring no expensive equipment or radioactivity.

Main Results:

  • The developed SH2 domain array assay is straightforward and cost-effective.
  • It enables the profiling of cellular phosphorylation status.
  • The assay is suitable for analyzing interactions under diverse biological conditions.

Conclusions:

  • The SH2 domain array provides a valuable tool for studying tyrosine phosphorylation and signal transduction.
  • This high-throughput method facilitates genome-wide phosphorylation profiling.
  • The assay's simplicity and lack of radioactivity make it broadly accessible.

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