SH2 domain specificity and activity modified by a single residue

Marengere, L. E. M.; Songyang, Zhou; Gish, Gerald; Schaller, Michael D.; Parsons, J. Thomas; Stern, Michael; Cantley, Lewis C.; Pawson, Tony

doi:10.1038/369502a0

Cited by 189 publications

(150 citation statements)

References 45 publications

(27 reference statements)

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“…The binding speci®city of the mutated domain correlated well with the biological activity as the mutated Src SH2 domain substituted the SH2 domain of the Grb2 protein in activation of the Ras pathway in vivo (Marengere et al, 1994). These two examples support the argument that there is a relatively simple relationship between the e determinants and binding speci®city.…”

Section: Sh2 Rulessupporting

confidence: 54%

From Src Homology domains to other signaling modules: proposal of the `protein recognition code'

1998

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The study of oncogenes has illuminated many aspects of cellular signaling. The delineation and characterization of protein modules exempli®ed by Src Homology domains has revolutionized our understanding of the molecular events underlying signal transduction pathways. Several well characterized intracellular modules which mediate protein-protein interactions, namely SH2, SH3, PH, PTB, EH, PDZ, EVH1 and WW domains, are directly involved in the multitude of membrane, cytoplasmic and nuclear processes in multicellular and/or unicellular organisms. The modular character of these protein domains and their cognate motifs, the universality of their molecular function, their widespread occurrence, and the speci®city as well as the degeneracy of their interactions have prompted us to propose the concept of the`protein recognition code'. By a parallel analogy to the universal genetic code, we propose here that there will be a ®nite set of precise rules to govern and predict protein-protein interactions mediated by modules. Several rules of the`protein recognition code' have already emerged.

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Section: Sh2 Rulessupporting

confidence: 54%

From Src Homology domains to other signaling modules: proposal of the `protein recognition code'

1998

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“…The burial of additional non-polar surface area upon ligand binding to monomeric Grb2-SH2 could explain its higher affinity for the latter. In this context, Marengere observed an approximately 14-fold reduction in the affinity for pTyr-Val-Asn-Val, a peptide similar to Ac-pYVN, when W121 was mutated to a threonine [36]. The crystallographic evidence presented herein clearly shows that the open conformation may be populated in ligand complexes.…”

Section: Discussionmentioning

confidence: 78%

Structural and energetic aspects of Grb2-SH2 domain-swapping

Benfield

Whiddon

Clements

et al. 2007

Archives of Biochemistry and Biophysics

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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.

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“…All known SH2 domains can be placed into four different groups on the basis of the amino acid at position ␤D5. The side chain at the ␤D5 position contacts pϩ1 and pϩ3 of the phospho-tyrosine ligand and has been shown to have a major effect on SH2 selectivity (3,24,25). In our set of control SH2 domains we included three closely related SH2 domains from signaling molecules Abl, Src, and Nck that are derived from the same group as Grb2 (group 1).…”

Section: Resultsmentioning

confidence: 99%

Specificity and affinity motifs for Grb2 SH2-ligand interactions

Kessels

Ward

Schumacher

2002

Proc. Natl. Acad. Sci. U.S.A.

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Protein-protein interactions are often mediated by the recognition of short continuous amino acid stretches on target proteins by specific binding domains. Affinity-based selection strategies have successfully been used to define recognition motifs for a large series of such protein domains. However, in many biological systems specificity of interaction may be of equal or greater importance than affinity. To address this issue we have developed a peptide library screening technology that can be used to directly define ligands for protein domains based on both affinity and specificity of interaction. We demonstrate the value of this approach by the selection of peptide ligands that are either highly specific for the Grb2 Src homology 2 (SH2) domain or that are cross-reactive between a group of related SH2 domains. Examination of previously identified physiological ligands for the Grb2 SH2 domain suggests that for these ligands regulation of the specificity of interaction may be an important factor for in vivo ligand selection.T he intracellular organization that enables a cell to respond to external stimuli consists of a complex web of signal transduction pathways. Key factors in the regulation of cellular signaling are the protein binding domains-small, conserved protein modules that mediate intracellular protein-protein interactions. Many of these domains, such as the families of SH2 (Src homology 2), SH3 (Src homology 3), PTB (phosphotyrosine binding), or PDZ (postsynaptic density-95͞Discs large͞zona occludens-1) domains, use short peptide sequences for ligand recognition (1). For example, the binding of SH2 domains to target proteins involves the recognition of a phosphorylated tyrosine residue, and specificity of individual SH2 domains is mediated by the recognition of amino acid residues immediately C-terminal to the phospho-tyrosine (2). The binding preferences of SH2 domains have been studied extensively through the use of peptide libraries, and predictions for the optimal binding motifs for a large number of SH2 domains have been obtained in this manner (3,4). In addition, such binding motifs have been used extensively as lead structures for the design of selective small-molecular SH2 inhibitors (5, 6). Importantly, in traditional library screening strategies used to define SH2 ligand motifs the selection of ligands is exclusively based on the strength of the SH2-phospho-peptide interaction. Consequently, the motifs that are identified in this manner describe ligands with an optimal affinity for a given SH2 domain (here named affinity motifs). Because both affinity and specificity of protein interactions are controlled by the same thermodynamic factors (shape and charge complementarity in the ground state), the selection of high affinity ligands will often also result in the selection of highly specific ligands. However, it has previously been argued that for closely related targets (such as the families of SH2 domains and other signal transduction modules) affinity-based selections may result in the ide...

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SH2 domain specificity and activity modified by a single residue

Cited by 189 publications

References 45 publications

From Src Homology domains to other signaling modules: proposal of the `protein recognition code'

From Src Homology domains to other signaling modules: proposal of the `protein recognition code'

Structural and energetic aspects of Grb2-SH2 domain-swapping

Specificity and affinity motifs for Grb2 SH2-ligand interactions

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