Related Experiment Video

Updated: Aug 18, 2026

Deciphering the Structural Effects of Activating EGFR Somatic Mutations with Molecular Dynamics Simulation
15:05

Deciphering the Structural Effects of Activating EGFR Somatic Mutations with Molecular Dynamics Simulation

Published on: May 20, 2020

Grb7-SH2 domain dimerisation is affected by a single point mutation

Corrine J Porter1, Matthew C J Wilce, Joel P Mackay

  • 1School of Biomedical and Chemical Sciences, University of Western Australia, 35 Stirling Highway, Crawley, Perth, WA 6009, Australia.

European Biophysics Journal : EBJ
|April 21, 2005
PubMed

Insights

Growth factor receptor bound protein 7 (Grb7) is an adaptor protein implicated in breast cancer. The Grb7 Src homology 2 (SH2) domain, crucial for ErbB2 interaction, was found to be dimeric, a property that can be disrupted by specific mutations.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cancer Research

Background:

  • Growth factor receptor bound protein 7 (Grb7) is an adaptor protein.
  • Grb7 is co-overexpressed with the ErbB2 receptor in breast tumors.
  • The interaction between Grb7 and ErbB2 is mediated by the Grb7 Src homology 2 (SH2) domain.

Purpose of the Study:

  • To investigate the self-association properties of the Grb7-SH2 domain.
  • To determine if the Grb7-SH2 domain exists as a monomer or dimer.
  • To identify mutations that alter the oligomeric state of the Grb7-SH2 domain for biophysical studies.

Main Methods:

  • Sedimentation equilibrium ultracentrifugation to assess protein oligomerization.
  • Size-exclusion chromatography to analyze the effect of mutations on protein structure.
  • Site-directed mutagenesis to alter specific amino acid residues.

Main Results:

  • The Grb7-SH2 domain was confirmed to exist as a homodimer in solution.
  • A dissociation constant (Kd) of approximately 11 μM was determined for the Grb7-SH2 domain dimer.
  • Mutation of phenylalanine 511 to arginine resulted in a monomeric form of the Grb7-SH2 domain.

Conclusions:

  • The Grb7-SH2 domain exhibits self-association, existing as a dimer.
  • Specific mutations, such as F511R, can disrupt dimerization and yield a monomeric Grb7-SH2 domain.
  • This engineered monomeric Grb7-SH2 domain is a valuable tool for future biophysical and structural investigations.

More Related Videos

Sedimentation Equilibrium of a Small Oligomer-forming Membrane Protein: Effect of Histidine Protonation on Pentameric Stability09:49

Sedimentation Equilibrium of a Small Oligomer-forming Membrane Protein: Effect of Histidine Protonation on Pentameric Stability

Published on: April 2, 2015

Related Concept Videos

Conserved Binding Sites01:49

Conserved Binding Sites

Many proteins’ biological role depends on their interactions with their ligands, small molecules that bind to specific locations on the protein known as ligand-binding sites. Ligand-binding sites are often conserved among homologous proteins as these sites are critical for protein function.
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally analyses the...
Point and Frameshift Mutations01:30

Point and Frameshift Mutations

Point mutations are genetic alterations involving the change of a single nucleotide base pair in DNA. Depending on how the alteration affects protein synthesis, they can lead to various consequences.Point mutations fall into the following types:Silent mutations occur when a nucleotide change does not alter the amino acid sequence due to the redundancy of the genetic code. For instance, changing ACC to ACA still encodes threonine, leaving the protein function unaffected. This occurs because...
Conservation of Protein Domains Over Different Proteins02:26

Conservation of Protein Domains Over Different Proteins

Protein domains are small structurally independent units that are part of a single amino acid chain.  Although these domains are often structurally independent, they may rely on synergistic effects to perform their functions as part of a larger protein. Protein domains may be conserved within the same organism, as well as across different organisms.
A limited set of protein domains often duplicate and recombine during evolution. These domains can be organized in different combinations to form...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
Mutations01:39

Mutations

Overview
Covalently Linked Protein Regulators02:04

Covalently Linked Protein Regulators

Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein.