Structural and Conformational Requirements for High-Affinity Binding to the SH2 Domain of Grb2

Ettmayer, Peter; Gounarides, John S.; Jarosinski, Mark A.; Martin, M.P.; Rondeau, Jean‐Michel; Sabio, Michael; Topiol, Sid; Weidmann, Beat; Zurini, Mauro; Bair, Kenneth W.

doi:10.1021/jm9811007

Cited by 87 publications

(102 citation statements)

References 31 publications

(51 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The significant differences between the complex of domain-swapped dimeric Grb2-SH2 with Ac-pYVN and the eight known crystal structures of monomericGrb2 SH2/ligand complexes [2,14,[32][33][34] are found in the conformations of residues 121-123 and 142 and their interactions with the domain and the side chain of the pY +2 Asn residue. In the domain-swapped dimer complex with Ac-pYVN, residues 121-123 are in an extended conformation that allows for formation of a hydrogen bond between the guanidino group of R142 and the carbonyl oxygen atom of V122.…”

Section: Discussionmentioning

confidence: 93%

Structural and energetic aspects of Grb2-SH2 domain-swapping

Benfield

Whiddon

Clements

et al. 2007

Archives of Biochemistry and Biophysics

View full text Add to dashboard Cite

The SH2 domain of growth factor receptor-bound protein 2 (Grb2) has been the focus of numerous studies, primarily because of the important roles it plays in signal transduction. More recently, it has emerged as a useful protein to study the consequences of ligand preorganization upon energetics and structure in protein-ligand interactions. The Grb2-SH2 domain is known to form a domain-swapped dimer, and as part of our investigations toward correlating structure and energetics in biological systems, we examined the effects that domain-swapping dimerization of the Grb2-SH2 domain had upon ligand binding affinities. Isothermal titration calorimetry was performed using Grb2-SH2 in both its monomeric and domain-swapped dimeric forms and a phosphorylated tripeptide AcNHpTyr-Val-Asn-NH 2 that is similar to the Shc sequence recognized by Grb2-SH2 in vivo. The two binding sites of domain-swapped dimer exhibited a 4-and a 13-fold reduction in ligand affinity compared to monomer. Crystal structures of peptide-bound and uncomplexed forms of Grb2-SH2 domain-swapped dimer were obtained and reveal that the orientation of residues V122, V123, and R142 may influence the conformation of W121, an amino acid that is believed to play an important role in Grb2-SH2 ligand sequence specificity. These findings suggest that domain-swapping of Grb2-SH2 not only results in a lower affinity for a Shc-derived ligand, but it may also affect ligand specificity.

show abstract

Section: Discussionmentioning

confidence: 93%

Structural and energetic aspects of Grb2-SH2 domain-swapping

Benfield

Whiddon

Clements

et al. 2007

Archives of Biochemistry and Biophysics

View full text Add to dashboard Cite

show abstract

“…33,40,41 From molecular modeling, it appeared that our cyclic compound (Chart 1) was able to adopt a similar -turn conformation as the bound linear ligand. However, the assumed favorable entropy due to preorganization in the bound conformation did not result in increased affinity.…”

Section: Discussionmentioning

confidence: 99%

Surface Plasmon Resonance Thermodynamic and Kinetic Analysis as a Strategic Tool in Drug Design. Distinct Ways for Phosphopeptides to Plug into Src- and Grb2 SH2 Domains

et al. 2005

View full text Add to dashboard Cite

Thermodynamic and kinetic studies of biomolecular interactions give insight into specificity of molecular recognition processes and advance rational drug design. Binding of phosphotyrosine (pY)-containing peptides to Src-and Grb2-SH2 domains was investigated using a surface plasmon resonance (SPR)-based method. This SPR assay yielded thermodynamic binding constants in solution, and the kinetic information contained in the SPR signal allowed kinetic analysis, which demonstrated distinct ways for pY ligands to interact with the SH2 domains. The results for binding to Src SH2 were consistent with sequestration of water molecules in the interface of the pYEEI peptide/Src SH2 complex. The results for a pYVNV peptide binding to Grb2 SH2 suggested a conformational change for Grb2 SH2 upon binding, which is not observed for Src SH2. Binding of a cyclic construct, allowing the pYVNV sequence in the bound conformation, did not have the expected entropy advantage. The results suggest an alternative binding mode for this construct, with the hydrophobic ring-closing part interacting with the protein. In all cases, except for full-length Grb2 protein, the affinity for the immobilized peptide at the SPR sensor and in solution was identical. This study demonstrates that SPR thermodynamic and kinetic analysis is a useful strategic tool in drug design.

show abstract

“…SH2 domains from the Grb2 family of proteins represent a distinct structural class of SH2 domain, due to the presence of a tryptophan residue (W121) in the EF loop which sterically hinders the binding of the pTyr ϩ 3 residue of the SH2 recognition motif (41,42). This feature imposes constraints on the conformation of phosphopeptide ligands for the SH2 domain which are quite different from those associated with SH2 domains from Src family kinases (40).…”

Section: Analysis Of the Grid Cd28 Interactionmentioning

confidence: 99%

GRID: A Novel Grb-2-Related Adapter Protein That Interacts with the Activated T Cell Costimulatory Receptor CD28

Ellis¹,

Ashman²,

Burden³

et al. 2000

The Journal of Immunology

View full text Add to dashboard Cite

Adapter proteins such as Grb2 play a central role in the formation of signaling complexes through their association with multiple protein binding partners. These interactions are mediated by specialized domains such as the well-characterized Src homology SH2 and SH3 motifs. Using yeast three-hybrid technology, we have identified a novel adapter protein, expressed predominantly in T lymphocytes, that associates with the activated form of the costimulatory receptor, CD28. The protein is a member of the Grb2 family of adapter proteins and contains an SH3-SH2-SH3 domain structure. A unique glutamine/proline-rich domain (insert domain) of unknown function is situated between the SH2 and N-terminal SH3 domains. We term this protein GRID for Grb2-related protein with insert domain. GRID coimmunoprecipitates with CD28 from Jurkat cell lysates following activation of CD28. Using mutants of CD28 and GRID, we demonstrate that interaction between the proteins is dependent on phosphorylation of CD28 at tyrosine 173 and integrity of the GRID SH2 domain, although there are also subsidiary stabilizing contacts between the PXXP motifs of CD28 and the GRID C-terminal SH3 domain. In addition to CD28, GRID interacts with a number of other T cell signaling proteins, including SLP-76 (SH2 domain-containing leukocyte protein of 76 kDa), p62dok, and RACK-1 (receptor for activated protein kinase C-1). These findings suggest that GRID functions as an adapter protein in the CD28-mediated costimulatory pathway in T cells.

show abstract

Structural and Conformational Requirements for High-Affinity Binding to the SH2 Domain of Grb2

Cited by 87 publications

References 31 publications

Structural and energetic aspects of Grb2-SH2 domain-swapping

Structural and energetic aspects of Grb2-SH2 domain-swapping

Surface Plasmon Resonance Thermodynamic and Kinetic Analysis as a Strategic Tool in Drug Design. Distinct Ways for Phosphopeptides to Plug into Src- and Grb2 SH2 Domains

GRID: A Novel Grb-2-Related Adapter Protein That Interacts with the Activated T Cell Costimulatory Receptor CD28

Contact Info

Product

Resources

About