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Related Experiment Videos

SH2 domain specificity and activity modified by a single residue

L E Marengere1, Z Songyang, G D Gish

  • 1Division of Molecular and Developmental Biology, Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada.

Nature
|June 9, 1994
PubMed
Summary

Researchers modified a key residue in the Src tyrosine kinase SH2 domain, switching its binding specificity. This engineered SH2 domain successfully activated the Ras pathway in vivo, demonstrating a link between SH2 binding specificity and biological function.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Cell Signaling

Background:

  • Protein-tyrosine kinases are crucial for cellular signaling.
  • SH2 domains mediate signal transduction by binding phosphotyrosine sites.
  • SH2 domain specificity is influenced by residues near the phosphotyrosine-binding pocket.

Purpose of the Study:

  • To investigate the role of specific residues in SH2 domain binding specificity.
  • To determine if altering SH2 domain specificity impacts biological activity.
  • To explore the functional consequences of modifying the Src SH2 domain.

Main Methods:

  • Site-directed mutagenesis of the Src SH2 domain (Thr to Trp substitution).
  • Assessing the binding specificity of the mutant Src SH2 domain.

Related Experiment Videos

  • In vivo functional assays using the Sem-5 protein and Ras pathway activation.

    • Main Results:

    • A single amino acid change (Thr to Trp) in the Src SH2 domain altered its binding specificity.
    • The mutant Src SH2 domain mimicked the specificity of the Sem-5/drk/Grb2 SH2 domain.
    • The engineered Src SH2 domain functionally replaced the Sem-5 SH2 domain in activating the Ras pathway in vivo.

    Conclusions:

    • Identified a critical residue that governs SH2 domain binding specificity.
    • Demonstrated that altering SH2 domain specificity can change its biological function.
    • Biological activity of SH2 domains is directly correlated with their binding specificity.