Src Homology Region 2 [SH2]-Domain Phosphatase or Src Homology Region 2 Domain-Containing PTP-1

A conserved noncatalytic domain SH2 (for src homology region 2) is located immediately N terminal to the kinase domains of all cytoplasmic protein-tyrosine kinases. We found that the wild-type v-fps SH2 domain stimulated the enzymatic activity of the adjacent kinase domain 10-fold and functioned as a powerful positive effector of catalytic and transforming activities within the v-fps oncoprotein (P130gag-fps). Partial proteolysis of P130gag-fps and supporting genetic data indicated that the v-fps SH2 domain exerts its effect on catalytic activity through an intramolecular interaction with the kinase domain. Amino acid alterations in the SH2 domain that impaired kinase function interfered with association of the SH2 domain with the kinase domain. Deletion of a conserved octapeptide motif converted the v-fps SH2 domain from an activator to an inhibitor of tyrosine kinase activity. This latent inhibitory activity of v-fps SH2 has functional implications for phospholipase C-gamma and p21ras GTPase-activating protein, both of which have two distinct SH2 domains suggestive of complex regulation. In addition to regulating the specific activity of the kinase domain, the SH2 domain of P130gag-fps was also found to be required for the tyrosine phosphorylation of specific cellular proteins, notably polypeptides of 124 and 62 kilodaltons. The SH2 domain therefore appears to play a dual role in regulation of kinase activity and recognition of cellular substrates.

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The common src homology region 2 domain of cytoplasmic signaling proteins is a positive effector of v-fps tyrosine kinase function

Molecular and Cellular Biology ◽

10.1128/mcb.9.10.4131-4140.1989 ◽

1989 ◽

Vol 9 (10) ◽

pp. 4131-4140

Author(s):

C A Koch ◽

M Moran ◽

I Sadowski ◽

T Pawson

Keyword(s):

Tyrosine Kinase ◽

Tyrosine Kinases ◽

Kinase Activity ◽

Specific Activity ◽

Intramolecular Interaction ◽

Kinase Domain ◽

Sh2 Domain ◽

Homology Region ◽

Src Homology ◽

Type V

A conserved noncatalytic domain SH2 (for src homology region 2) is located immediately N terminal to the kinase domains of all cytoplasmic protein-tyrosine kinases. We found that the wild-type v-fps SH2 domain stimulated the enzymatic activity of the adjacent kinase domain 10-fold and functioned as a powerful positive effector of catalytic and transforming activities within the v-fps oncoprotein (P130gag-fps). Partial proteolysis of P130gag-fps and supporting genetic data indicated that the v-fps SH2 domain exerts its effect on catalytic activity through an intramolecular interaction with the kinase domain. Amino acid alterations in the SH2 domain that impaired kinase function interfered with association of the SH2 domain with the kinase domain. Deletion of a conserved octapeptide motif converted the v-fps SH2 domain from an activator to an inhibitor of tyrosine kinase activity. This latent inhibitory activity of v-fps SH2 has functional implications for phospholipase C-gamma and p21ras GTPase-activating protein, both of which have two distinct SH2 domains suggestive of complex regulation. In addition to regulating the specific activity of the kinase domain, the SH2 domain of P130gag-fps was also found to be required for the tyrosine phosphorylation of specific cellular proteins, notably polypeptides of 124 and 62 kilodaltons. The SH2 domain therefore appears to play a dual role in regulation of kinase activity and recognition of cellular substrates.

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Src Homology Region 2 (SH2)-Domain Phosphatase or Src Homology Region 2 Domain-Containing PTP-1 (SHP-1 or SH-PTP1)

Encyclopedia of Signaling Molecules ◽

10.1007/978-1-4419-0461-4_101271 ◽

2012 ◽

pp. 1779-1779

Author(s):

Brian R. Dempsey ◽

Anne C. Rintala-Dempsey ◽

Gary S. Shaw ◽

Yuan Xiao Zhu ◽

A. Keith Stewart ◽

...

Keyword(s):

Sh2 Domain ◽

Homology Region ◽

Src Homology

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Design Potential Selective Inhibitors for Treating Cancer by Targeting the Src Homology 2 (SH2) Domain-Containing Phosphatase 2 (Shp2) with Core Hopping Approach

Protein and Peptide Letters ◽

10.2174/0929866521666131223143913 ◽

2014 ◽

Vol 21 (6) ◽

pp. 556-563 ◽

Cited By ~ 10

Author(s):

Yu-Qing Duan ◽

Ying Ma ◽

Xue-Jiao Wang ◽

Yuan-Yuan Jin ◽

Run-Ling Wang ◽

...

Keyword(s):

Sh2 Domain ◽

Selective Inhibitors ◽

Src Homology 2 ◽

Src Homology

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PLC-γ directly binds activated c-Src, which is necessary for carbachol-mediated inhibition of NHE3 activity in Caco-2/BBe cells

AJP Cell Physiology ◽

10.1152/ajpcell.00277.2012 ◽

2013 ◽

Vol 305 (3) ◽

pp. C266-C275 ◽

Cited By ~ 5

Author(s):

Nicholas C. Zachos ◽

Luke J. Lee ◽

Olga Kovbasnjuk ◽

Xuhang Li ◽

Mark Donowitz

Keyword(s):

Sh2 Domain ◽

Signaling Proteins ◽

Multiprotein Complexes ◽

Sh2 Domains ◽

Src Homology 2 ◽

Src Inhibitor ◽

Intracellular Ca ◽

Direct Binding ◽

Src Homology

Elevated levels of intracellular Ca2+([Ca2+]i) inhibit Na+/H+exchanger 3 (NHE3) activity in the intact intestine. We previously demonstrated that PLC-γ directly binds NHE3, an interaction that is necessary for [Ca2+]iinhibition of NHE3 activity, and that PLC-γ Src homology 2 (SH2) domains may scaffold Ca2+signaling proteins necessary for regulation of NHE3 activity. [Ca2+]iregulation of NHE3 activity is also c-Src dependent; however, the mechanism by which c-Src is involved is undetermined. We hypothesized that the SH2 domains of PLC-γ might link c-Src to NHE3-containing complexes to mediate [Ca2+]iinhibition of NHE3 activity. In Caco-2/BBe cells, carbachol (CCh) decreased NHE3 activity by ∼40%, an effect abolished with the c-Src inhibitor PP2. CCh treatment increased the amount of active c-Src as early as 1 min through increased Y416phosphorylation. Coimmunoprecipitation demonstrated that c-Src associated with PLC-γ, but not NHE3, under basal conditions, an interaction that increased rapidly after CCh treatment and occurred before the dissociation of PLC-γ and NHE3 that occurred 10 min after CCh treatment. Finally, direct binding to c-Src only occurred through the PLC-γ SH2 domains, an interaction that was prevented by blocking the PLC-γ SH2 domain. This study demonstrated that c-Src 1) activity is necessary for [Ca2+]iinhibition of NHE3 activity, 2) activation occurs rapidly (∼1 min) after CCh treatment, 3) directly binds PLC-γ SH2 domains and associates dynamically with PLC-γ under elevated [Ca2+]iconditions, and 4) does not directly bind NHE3. Under elevated [Ca2+]iconditions, PLC-γ scaffolds c-Src into NHE3-containing multiprotein complexes before dissociation of PLC-γ from NHE3 and subsequent endocytosis of NHE3.

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Synthesis and in Vitro Evaluation of a Peptidomimetic Inhibitor Targeting the Src Homology 2 (SH2) Domain of STAT6

ACS Medicinal Chemistry Letters ◽

10.1021/ml4003919 ◽

2013 ◽

Vol 5 (1) ◽

pp. 69-72 ◽

Cited By ~ 15

Author(s):

Pietro Morlacchi ◽

Pijus K. Mandal ◽

John S. McMurray

Keyword(s):

Sh2 Domain ◽

In Vitro Evaluation ◽

Src Homology 2 ◽

Src Homology

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Enhanced Prediction of Src Homology 2 (SH2) Domain Binding Potentials Using a Fluorescence Polarization-derived c-Met, c-Kit, ErbB, and Androgen Receptor Interactome

Molecular & Cellular Proteomics ◽

10.1074/mcp.m113.034876 ◽

2014 ◽

Vol 13 (7) ◽

pp. 1705-1723 ◽

Cited By ~ 9

Author(s):

Kin K. Leung ◽

Ronald J. Hause ◽

John L. Barkinge ◽

Mark F. Ciaccio ◽

Chih-Pin Chuu ◽

...

Keyword(s):

Androgen Receptor ◽

Fluorescence Polarization ◽

Sh2 Domain ◽

Src Homology 2 ◽

Src Homology

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Evidence for a molecular complex consisting of Fyb/SLAP, SLP-76, Nck, VASP and WASP that links the actin cytoskeleton to Fcγ receptor signalling during phagocytosis

Journal of Cell Science ◽

10.1242/jcs.114.23.4307 ◽

2001 ◽

Vol 114 (23) ◽

pp. 4307-4318

Author(s):

Marc G. Coppolino ◽

Matthias Krause ◽

Petra Hagendorff ◽

David A. Monner ◽

William Trimble ◽

...

Keyword(s):

Actin Cytoskeleton ◽

Molecular Complex ◽

Actin Filaments ◽

Sh2 Domain ◽

Fcγ Receptors ◽

Receptor Signalling ◽

Src Homology 2 ◽

Src Homology ◽

Particle Ingestion ◽

Syndrome Protein

Phagocytosis by macrophages and neutrophils involves the spatial and temporal reorganisation of the actin-based cytoskeleton at sites of particle ingestion. Local polymerisation of actin filaments supports the protrusion of pseudopodia that eventually engulf the particle. Here we have investigated in detail the cytoskeletal events initiated upon engagement of Fc receptors in macrophages. Ena/vasodilator-stimulated phosphoprotein (VASP) proteins were recruited to phagosomes forming around opsonised particles in both primary and immortalised macrophages. Not only did the localisation of Ena/VASP proteins coincide, spatially and temporally, with the phagocytosis-induced reorganisation of actin filaments, but their recruitment to the phagocytic cup was required for the remodelling of the actin cytoskeleton, extension of pseudopodia and efficient particle internalisation. We also report that SLP-76, Vav and profilin were recruited to forming phagosomes. Upon induction of phagocytosis, a large molecular complex, consisting in part of Ena/VASP proteins, the Fyn-binding/SLP-76-associated protein (Fyb/SLAP), Src-homology-2 (SH2)-domain-containing leukocyte protein of 76 kDa (SLP-76), Nck, and the Wiskott-Aldrich syndrome protein (WASP), was formed. Our findings suggest that activation of Fcγ receptors triggers two signalling events during phagocytosis: one through Fyb/SLAP that leads to recruitment of VASP and profilin; and another through Nck that promotes the recruitment of WASP. These converge to regulate actin polymerisation, controlling the assembly of actin structures that are essential for the process of phagocytosis.

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Src Homology 2 (SH2)-Containing 5′-Inositol Phosphatase Localizes to Podosomes, and the SH2 Domain Is Implicated in the Attenuation of Bone Resorption in Osteoclasts

Endocrinology ◽

10.1210/en.2005-1309 ◽

2006 ◽

Vol 147 (7) ◽

pp. 3307-3317 ◽

Cited By ~ 19

Author(s):

Keiichiro Yogo ◽

Megumi Mizutamari ◽

Kazuta Mishima ◽

Hiromi Takenouchi ◽

Norihiro Ishida-Kitagawa ◽

...

Keyword(s):

Bone Resorption ◽

Nuclear Factor Κb ◽

Sh2 Domain ◽

Terminal Region ◽

Spectrometric Analysis ◽

Inositol Phosphatase ◽

Amino Terminal ◽

Novel Proteins ◽

Src Homology 2 ◽

Src Homology

c-Src plays an important role in bone resorption by osteoclasts. Here, we show using wild-type and ship−/− osteoclasts that Src homology 2 (SH2)-containing 5′-inositol phosphatase (SHIP) appeared to negatively regulate bone resorption activated by c-Src. SHIP was found to localize to podosomes under the influence of c-Src, and the presence of either the amino-terminal region comprising the SH2 domain or the carboxyl-terminal region was sufficient for its localization. Although SHIP lacking a functional SH2 domain was still found in podosomes, it could not rescue the hyper-bone resorbing activity and hypersensitivity to receptor activator of nuclear factor-κB ligand in ship−/− osteoclasts, suggesting that the localization of SHIP to podosomes per se was not sufficient and the SH2 domain was indispensable for its function. Cas and c-Cbl, known to function in podosomes of osteoclasts, were identified as novel proteins binding to the SHIP SH2 domain by mass spectrometric analysis, and this interaction appeared to be dependent on the Src kinase activity. These results demonstrate that c-Src enhances the translocation of SHIP to podosomes and regulates its function there through the SH2 domain, leading to an attenuation of bone resorption.

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