Structural basis for specificity of GRB2-SH2 revealed by a novel ligand binding mode

Rahuel, Joseph; Erdmann, Dirk; Strauss, André; García‐Echeverría, Carlos; Furet, Pascal; Caravatti, Giorgio; Fretz, Heinz; Schoepfer, Joseph; Grütter, Markus G.

doi:10.1038/nsb0796-586

Cited by 239 publications

(289 citation statements)

References 34 publications

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“…6). This structural homology is particularly suggestive in view of the fact that phospho-peptide ligands can bind only the Grb2 SH2 domain in a unique ␤-turn conformation, which is not observed in phospho-peptide ligands for other SH2 domains (37,38). However, the assignment of these YKNI loops within SHP-1 and SHP-2 as tyrosine phosphorylation sites and as putative Grb2 binding sites remains hypothetical in the absence of direct experimental confirmation.…”

Section: Resultsmentioning

confidence: 96%

“…This observation is consistent with sequence and structural analysis of SH2 domains. Accommodation of a lysine at pϩ1 in a phospho-peptide structure that binds Grb2 SH2 may be facilitated by interaction of the lysine -amino group with Gln-106 (␤D-strand) within the binding pocket (37). In contrast, lysine side chains may be excluded at pϩ1 for ligands of the C-terminal and N-terminal SH2 domains of p85 because of 7.…”

Section: Discussionmentioning

confidence: 99%

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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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Section: Resultsmentioning

confidence: 96%

Section: Discussionmentioning

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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“…Fig. 3b shows this segment for c-Met in comparison to the same sequence of a phosphopeptide bound to Grb2 SH2 (39). Conformational differences concern the -dihedral angle of Tyr-1356 and its side chain rotamer, as well as a peptide flip for the bond between Val-1357 and Asn-1358.…”

Section: Resultsmentioning

confidence: 99%

“…In general, phosphotyrosine-containing peptides bind to SH2 domains in an extended conformation (38). Grb2 SH2 is the only SH2 domain reported to date that binds the peptides in a type I ␤-turn conformation with the asparagine residue in the position pY ϩ 2 forming two hydrogen bonds with the SH2 domain (39,40). The bulky Trp-121 side chain in the EF loop of Grb2-SH2 excludes an extended peptide conformation.…”

Section: Resultsmentioning

confidence: 99%

Crystal structure of the tyrosine kinase domain of the hepatocyte growth factor receptor c-Met and its complex with the microbial alkaloid K-252a

Schiering

Knapp²,

Marconi³

et al. 2003

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

140

133

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“…Moreover, the Shc SH2 domain is monomeric in solution at physiologic pH (37). A domain-swapped dimeric form of the Grb2 (unrelated to Grb7/Grb10/Grb14) SH2 domain has been observed crystallographically (38), as has a monomeric form (39), but this dimer is likely to be an artifact of expression as a glutathione S-transferase fusion protein (40).…”

Section: Discussionmentioning

confidence: 99%

Structural Basis for Dimerization of the Grb10 Src Homology 2 Domain

Stein

Ghirlando

Hubbard

2003

Journal of Biological Chemistry

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Grb7, Grb10, and Grb14 are members of a distinct family of adapter proteins that interact with various receptor tyrosine kinases upon receptor activation. Proteins in this family contain several modular signaling domains including a pleckstrin homology (PH) domain, a BPS (between PH and SH2) domain, and a C-terminal Src homology 2 (SH2) domain. Although SH2 domains are typically monomeric, we show that the Grb10 SH2 domain and also full-length Grb10␥ are dimeric in solution under physiologic conditions. The crystal structure of the Grb10 SH2 domain at 1.65-Å resolution reveals a non-covalent dimer whose interface comprises residues within and flanking the C-terminal ␣ helix, which are conserved in the Grb7/ Grb10/Grb14 family but not in other SH2 domains. Val-522 in the BG loop (BG3) and Asp-500 in the EF loop (EF1) are positioned to interfere with the binding of the P؉3 residue of a phosphopeptide ligand. These structural features of the Grb10 SH2 domain will favor binding of dimeric, turn-containing phosphotyrosine sequences, such as the phosphorylated activation loops in the two ␤ subunits of the insulin and insulin-like growth factor-1 receptors. Moreover, the structure suggests the mechanism by which the Grb7 SH2 domain binds selectively to pTyr-1139 (pYVNQ) in Her2, which along with Grb7 is co-amplified in human breast cancers.Grb10 is a member of a family of adapter proteins including Grb7 and Grb14, which have been identified as putative downstream effectors of receptor tyrosine kinases (1). Proteins in this family contain several modular domains including an Nterminal proline-rich region, a Ras-associated-like domain, a pleckstrin homology (PH) 1 domain, a short region known as the BPS (between PH and SH2) or PIR (phosphorylated insulin receptor-interacting region) domain, and a C-terminal Src homology 2 (SH2) domain. Grb10 has multiple alternatively spliced forms (␣-), which differ in their N-terminal region including the PH domain.The BPS/PIR and SH2 domains of Grb7/Grb10/Grb14 have been implicated in the interaction with activated receptor tyrosine kinases. The BPS domain is a structurally uncharacterized region of ϳ50 residues that is unique to this family of adapter proteins. The SH2 domain, found in many signaling proteins, is a well characterized protein module of ϳ100 residues that binds phosphotyrosine-containing sequences (2, 3). The BPS domains of Grb7/Grb10/Grb14 bind to the phosphorylated insulin and insulin-like growth factor-1 (IGF1) receptors (4 -6). The SH2 domains of Grb7/Grb10/Grb14 also interact with the insulin and IGF1 receptors (4, 5, 7-11) and, in addition, have been shown to mediate the interaction with the epidermal growth factor receptor (4, 12), Her2 (epidermal growth factor receptor family) (13, 14), platelet-derived growth factor receptor-␤ (10, 15, 16), Ret (17), EphB1 (18), Kit (19), Tie2 (20), and fibroblast growth factor receptor-1 (21).The roles of Grb10 family members in growth factor-mediated signaling have not been clearly established. Most studies have investigated...

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Structural basis for specificity of GRB2-SH2 revealed by a novel ligand binding mode

Cited by 239 publications

References 34 publications

Specificity and affinity motifs for Grb2 SH2-ligand interactions

Specificity and affinity motifs for Grb2 SH2-ligand interactions

Crystal structure of the tyrosine kinase domain of the hepatocyte growth factor receptor c-Met and its complex with the microbial alkaloid K-252a

Structural Basis for Dimerization of the Grb10 Src Homology 2 Domain

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