SH2 Domain Structures and Interactions

Gopalasingam, Piraveen; Quill, Lee; Jeeves, Mark; Overduin, Michael

doi:10.1007/978-3-319-20098-9_8

Cited by 7 publications

(10 citation statements)

References 103 publications

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“…Tyrosine phosphorylation contributes to only ~0.5% of the total phosphoproteome, yet it plays critical roles in eukaryotic cell regulation [Gopalasingam 2015]. Substrate specificities of kinases and phosphatases are broad, and their effects in signaling are controlled also by their location.…”

Section: Sh2 Domainsmentioning

confidence: 99%

“…300 three-dimensional structures of approximately 70 different SH2 domains have been determined. They reveal a highly conserved topology [Gopalasingam 2015;Liu 2017]. These domains contain approximately 100 amino acids, with a central β strand, flanked by two α helices.…”

Section: Structure Of the Sh2 Domainsmentioning

confidence: 99%

“…Association of the SH2 domains to pY motifs favors N-SH2/PTP dissociation and thereby activation of the phosphatase ( Figure 1D) [Gopalasingam 2015]. The loss of the N-SH2/PTP interactions is triggered by a conformational transition of N-SH2 that leads to a loss of complementarity between the N-SH2 and PTP surfaces.…”

Section: The Sh2 Domain-containing Protein Tyrosine Phosphatasementioning

confidence: 99%

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Structural Determinants of Phosphopeptide Binding to the N-Terminal Src Homology 2 Domain of the SHP2 Phosphatase

Anselmi

Calligari

Hub

et al. 2020

Preprint

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ABSTRACTSH2 domain-containing tyrosine phosphatase 2 (SHP2), encoded by PTPN11, plays a fundamental role in the modulation of several signaling pathways. Germline and somatic mutations in PTPN11 are associated with different rare diseases and hematologic malignancies, and recent studies have individuated SHP2 as a central node in oncogenesis and cancer drug resistance. SHP2 structure includes two Src homology 2 domains (N-SH2 and C-SH2) followed by a catalytic protein tyrosine phosphatase (PTP) domain. Under basal conditions, the N-SH2 domain blocks the active site, inhibiting phosphatase activity. Association of the N-SH2 domain with binding partners containing short amino acid motifs comprising a phosphotyrosine residue (pY) leads to N-SH2/PTP dissociation and SHP2 activation. Considering the relevance of SHP2 in signaling and disease and the central role of the N-SH2 domain in its allosteric regulation mechanism, we performed microsecond-long molecular dynamics simulations of the N-SH2 domain complexed to 12 different peptides, to define the structural and dynamical features determining the binding affinity and specificity of the domain. Phosphopeptide residues at position −2 to +5, with respect to pY, have significant interactions with the SH2 domain. In addition to the strong interaction of the pY residue with its conserved binding pocket, the complex is stabilized hydrophobically by insertion of residues +1, +3 and +5 in an apolar groove of the domain, and interaction of residue −2 with both the pY and a protein surface residue. Additional interactions are provided by hydrogen bonds formed by the backbone of residues −1, +1, +2 and +4. Finally, negatively charged residues at position +2 and +4 are involved in electrostatic interactions with two lysines (Lys89 and Lys91) specific of the SHP2 N-SH2 domain. Interestingly, the MD simulations illustrated a previously undescribed conformational flexibility of the domain, involving the core β-sheet and the loop that closes the pY binding pocket.

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Section: Sh2 Domainsmentioning

confidence: 99%

Section: Structure Of the Sh2 Domainsmentioning

confidence: 99%

Section: The Sh2 Domain-containing Protein Tyrosine Phosphatasementioning

confidence: 99%

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Structural Determinants of Phosphopeptide Binding to the N-Terminal Src Homology 2 Domain of the SHP2 Phosphatase

Anselmi

Calligari

Hub

et al. 2020

Preprint

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“…18 The structural studies show that the Y/F substitutions would eliminate critical contacts that allow the deep simultaneous insertion of G6b-B's ITIM and ITSM phosphotyrosine residues into Shp2 N-SH2 and C-SH2 domains, respectively, in a manner that is consistent with canonical SH2 ligand binding modes. 19 Moreover, the interaction with Shp2 tandem SH2 domains would also be compromised by G6b-B deletions and point mutations in Shp2, including L210I, R223H/C, and A238V, which have been found in human tumors, [20][21][22] suggesting wider roles for Shp2 uncoupling in cancer.…”

Section: Shp2 Interacts Directly With Phosphorylated G6b-b In An Extementioning

confidence: 99%

Uncoupling ITIM receptor G6b-B from tyrosine phosphatases Shp1 and Shp2 disrupts murine platelet homeostasis

Geer

Geffen

Gopalasingam

et al. 2018

Blood

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The immunoreceptor tyrosine-based inhibitory motif (ITIM)-containing receptor G6b-B has emerged as a key regulator of platelet homeostasis. However, it remains unclear how it mediates its effects. Tyrosine phosphorylation of ITIM and immunoreceptor tyrosine-based switch motif (ITSM) within the cytoplasmic tail of G6b-B provides a docking site for Src homology 2 domain-containing protein-tyrosine phosphatases Shp1 and Shp2, which are also critical regulators of platelet production and function. In this study, we investigate the physiological consequences of uncoupling G6b-B from Shp1 and Shp2. To address this, we generated a transgenic mouse model expressing a mutant form of G6b-B in which tyrosine residues 212 and 238 within ITIM and ITSM were mutated to phenylalanine. Mice homozygous for the mutation () were macrothrombocytopenic, as a result of the reduction in platelet production, and had large clusters of megakaryocytes and myelofibrosis at sites of hematopoiesis, similar to those observed in -deficient mice and patients. Platelets from mice were hyporesponsive to collagen, as a result of the significant reduction in the expression of the immunoreceptor tyrosine-based activation motif (ITAM)-containing collagen receptor complex GPVI-FcR γ-chain, as well as thrombin, which could be partially rescued by costimulating the platelets with adenosine diphosphate. In contrast, platelets from ,, and megakaryocyte-specific mice were hyperresponsive to antibody-mediated cross-linking of the hemi-ITAM-containing podoplanin receptor CLEC-2, suggesting that G6b-B inhibits CLEC-2-mediated platelet activation through Shp2. Findings from this study demonstrate that G6b-B must engage with Shp1 and Shp2 to mediate its regulatory effects on platelet homeostasis.

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“…Such moderate affinities are considered to be crucial for allowing transient association and dissociation events in cell signaling. Consistently, SH2 domains artificially engineered to reach low nanomolar affinities for phosphorylated sequences (known as superbinders) [Kaneko 2012 b], by increasing the domain affinity for the pY residue, have detrimental consequences for signal transduction [Gopalasingam 2015].…”

Section: Discussionmentioning

confidence: 99%

Targeting Oncogenic Src Homology 2 Domain-Containing Phosphatase 2 (SHP2) by Inhibiting its Protein-Protein Interactions

Bobone

Pannone

Biondi

et al. 2020

Preprint

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Mutations of PTPN11, the gene coding for the Src homology 2 domain-containing phosphatase 2 (SHP2), cause childhood leukemias and developmental disorders. SHP2 inhibitors targeting the catalytic site or an allosteric pocket lack specificity or are ineffective on pathogenic variants. In addition, several data indicate that increased association with cognate proteins, through its SH2 domains, rather than enhanced catalytic activity, is the main effect of mutations causing hyperactivation of SHP2-mediated signaling. We developed peptide-based molecules with low nM affinity to the N-SH2 domain and high specificity. These molecules bind to pathogenic variants of SHP2 with an affinity up to 20 times higher than to the wild-type protein, in contrast to allosteric inhibitors, and were able to revert the effects of a pathogenic SHP2 mutation in zebrafish embryos. Our results provide a novel route for SHP2-targeted therapies and a tool to investigate the role of protein-protein interactions in the function of SHP2.TABLE OF CONTENTS GRAPHICS

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SH2 Domain Structures and Interactions

Cited by 7 publications

References 103 publications

Structural Determinants of Phosphopeptide Binding to the N-Terminal Src Homology 2 Domain of the SHP2 Phosphatase

Structural Determinants of Phosphopeptide Binding to the N-Terminal Src Homology 2 Domain of the SHP2 Phosphatase

Uncoupling ITIM receptor G6b-B from tyrosine phosphatases Shp1 and Shp2 disrupts murine platelet homeostasis

Targeting Oncogenic Src Homology 2 Domain-Containing Phosphatase 2 (SHP2) by Inhibiting its Protein-Protein Interactions

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