A Drosophila shc gene product is implicated in signaling by the DER receptor tyrosine kinase.

Antibodies to the human Shc adaptor protein were used to isolate a cDNA encoding a Drosophila Shc protein (dShc) by screening an expression library. The dshc gene, which maps to position 67B-C on the third chromosome, encodes a 45-kDa protein that is widely expressed throughout the Drosophila life cycle. In flies, the dShc protein physically associates with the activated Drosophila epidermal growth factor receptor homolog (DER) and is inducibly phosphorylated on tyrosine by DER. The 45-kDa dShc protein is closely related both in overall organization and in amino acid sequence (46% identity) to the 52-kDa mammalian Shc isoform. In addition to a C-terminal Src homology 2 (SH2) domain, dShc contains an N-terminal phosphotyrosine-binding (PTB) domain, which associates in vitro with the autophosphorylated DER receptor tyrosine kinase and with phosphopeptides containing an Asn-Pro-X-pTyr motif, where pTyr stands for phosphotyrosine. A potential binding site for the dShc PTB domain is located at Tyr-1228 of DER. These results indicate that the shc gene has been conserved in evolution, as have the binding properties of the Shc PTB and SH2 domains. Despite the close relationship between the Drosophila and mammalian Shc proteins, dShc lacks the high-affinity Grb2-binding site found in mammalian Shc, suggesting that Shc proteins may have functions in addition to regulation of the Ras pathway.

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Disabled Is a Putative Adaptor Protein That Functions during Signaling by the Sevenless Receptor Tyrosine Kinase

Molecular and Cellular Biology ◽

10.1128/mcb.18.8.4844 ◽

1998 ◽

Vol 18 (8) ◽

pp. 4844-4854 ◽

Cited By ~ 17

Author(s):

Ngocdiep Le ◽

Michael A. Simon

Keyword(s):

Tyrosine Kinase ◽

Signaling Pathway ◽

Receptor Tyrosine Kinase ◽

Biochemical Analysis ◽

Adaptor Protein ◽

Sh2 Domain ◽

Sh3 Domains ◽

Sevenless Receptor Tyrosine Kinase ◽

Ptb Domain

ABSTRACT DRK, the Drosophila homolog of the SH2-SH3 domain adaptor protein Grb2, is required during signaling by thesevenless receptor tyrosine kinase (SEV). One role of DRK is to provide a link between activated SEV and the Ras1 activator SOS. We have investigated the possibility that DRK performs other functions by identifying additional DRK-binding proteins. We show that the phosphotyrosine-binding (PTB) domain-containing protein Disabled (DAB) binds to the DRK SH3 domains. DAB is expressed in the ommatidial clusters, and loss of DAB function disrupts ommatidial development. Moreover, reduction of DAB function attenuates signaling by a constitutively activated SEV. Our biochemical analysis suggests that DAB binds SEV directly via its PTB domain, becomes tyrosine phosphorylated upon SEV activation, and then serves as an adaptor protein for SH2 domain-containing proteins. Taken together, these results indicate that DAB is a novel component of the SEV signaling pathway.

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The Selectivity of Receptor Tyrosine Kinase Signaling Is Controlled by a Secondary SH2 Domain Binding Site

Cell ◽

10.1016/j.cell.2009.05.028 ◽

2009 ◽

Vol 138 (3) ◽

pp. 514-524 ◽

Cited By ~ 95

Author(s):

Jae Hyun Bae ◽

Erin Denise Lew ◽

Satoru Yuzawa ◽

Francisco Tomé ◽

Irit Lax ◽

...

Keyword(s):

Tyrosine Kinase ◽

Binding Site ◽

Receptor Tyrosine Kinase ◽

Sh2 Domain ◽

Kinase Signaling ◽

Tyrosine Kinase Signaling

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The Shc adaptor protein forms interdependent phosphotyrosine-mediated protein complexes in mast cells stimulated with interleukin 3

Blood ◽

10.1182/blood.v96.1.132.013k13_132_138 ◽

2000 ◽

Vol 96 (1) ◽

pp. 132-138 ◽

Cited By ~ 1

Author(s):

Laura Velazquez ◽

Gerald D. Gish ◽

Peter van der Geer ◽

Lorne Taylor ◽

Johanna Shulman ◽

...

Keyword(s):

Mast Cells ◽

Protein Complexes ◽

Adaptor Protein ◽

Phospholipid Metabolism ◽

Sh2 Domain ◽

Interleukin 3 ◽

Inositol Phosphatase ◽

Ptb Domain

The Shc adaptor protein possesses 2 distinct phosphotyrosine (pTyr) recognition modules—the pTyr binding (PTB) domain and the Src homology 2 (SH2) domain—and multiple potential sites for tyrosine (Tyr) phosphorylation (Tyr residues 239, 240, and 317). On stimulation of hematopoietic cells with interleukin 3 (IL-3), Shc becomes phosphorylated and may therefore contribute to IL-3 signaling. We investigated the interactions mediated by the Shc modular domains and pTyr sites in IL-3–dependent IC2 premast cells. The Shc PTB domain, rather than the SH2 domain, associated both in vitro and in vivo with the Tyr-phosphorylated β subunit of the IL-3 receptor and with the SH2-containing 5′ inositol phosphatase (SHIP), and it recognized specific NXXpY phosphopeptides from these binding partners. In IL-3–stimulated mast cells, Shc phosphorylation occurred primarily on Tyr239 and 317 and was dependent on a functional PTB domain. Phosphorylated Tyr317, and to a lesser extent, Tyr239, bound the Grb2 adaptor and SHIP. Furthermore, a pTyr317 Shc phosphopeptide selectively recognized Grb2, Sos1, SHIP, and the p85 subunit of phosphatidylinositol 3′ kinase from mast cells, as characterized by mass spectrometry. These results indicate that Shc undergoes an interdependent series of pTyr-mediated interactions in IL-3–stimulated mast cells, resulting in the recruitment of proteins that regulate the Ras pathway and phospholipid metabolism.

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Exploring the interaction between epidermal growth factor receptor tyrosine kinase and some of the synthesized inhibitors using combination of in-silico and in-vitro cytotoxicity methods

Research in Pharmaceutical Sciences ◽

10.4103/1735-5362.245963 ◽

2018 ◽

Vol 13 (6) ◽

pp. 509 ◽

Cited By ~ 4

Author(s):

Mahboubeh Mansourian ◽

RezvanRezaee Nasab ◽

Farshid Hassanzadeh ◽

Mohsen Shahlaei

Keyword(s):

Epidermal Growth Factor Receptor ◽

Growth Factor ◽

Epidermal Growth Factor ◽

Tyrosine Kinase ◽

Receptor Tyrosine Kinase ◽

In Silico ◽

Growth Factor Receptor ◽

In Vitro Cytotoxicity ◽

Epidermal Growth

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In Vitro and In Silico Studies of Quercetin and Daidzin as Selective Anticancer Agents

Indonesian Journal of Chemistry ◽

10.22146/ijc.53552 ◽

2021 ◽

Vol 21 (2) ◽

pp. 310

Author(s):

Muhammad Sulaiman Zubair ◽

Syariful Anam ◽

Saipul Maulana ◽

Muhammad Arba

Keyword(s):

Epidermal Growth Factor Receptor ◽

Molecular Docking ◽

Growth Factor ◽

Epidermal Growth Factor ◽

Tyrosine Kinase ◽

Receptor Tyrosine Kinase ◽

Growth Factor Receptor ◽

In Silico Studies ◽

Epidermal Growth

Quercetin and daidzin are flavonoid and flavonoid glycoside type compounds that have been found in many plants and nutraceuticals. This study aims to examine the in vitro cytotoxic and selectivity properties of quercetin and daidzin on breast and cervical cancers and to study their molecular interaction and stability on epidermal growth factor receptor tyrosine kinase (EGFR-TK) by applying molecular docking and molecular dynamics (MD) simulations. In vitro anticancer activity was performed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method on breast cancer cell (T47D), cervical cancer cells (HeLa), and Vero normal cells, while molecular docking and MD simulation were done by using AutoDock Vina and Amber18 package software, respectively. Quercetin and daidzin showed potent cytotoxic and high selectivity on both cell lines. Daidzin was found to has a higher IC50 and selectivity index than quercetin. Docking and MD results showed that both compounds prefer to interact with epidermal growth factor receptor tyrosine kinase (EGFR-TK). Daidzin showed better interaction than quercetin with a docking score of -9.6 kcal/mol. Also, daidzin was found more stable than quercetin with low RMSD and RMSF values.

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The Shc adaptor protein forms interdependent phosphotyrosine-mediated protein complexes in mast cells stimulated with interleukin 3

Blood ◽

10.1182/blood.v96.1.132 ◽

2000 ◽

Vol 96 (1) ◽

pp. 132-138 ◽

Cited By ~ 16

Author(s):

Laura Velazquez ◽

Gerald D. Gish ◽

Peter van der Geer ◽

Lorne Taylor ◽

Johanna Shulman ◽

...

Keyword(s):

Mast Cells ◽

Protein Complexes ◽

Adaptor Protein ◽

Phospholipid Metabolism ◽

Sh2 Domain ◽

Interleukin 3 ◽

Inositol Phosphatase ◽

Ptb Domain

Abstract The Shc adaptor protein possesses 2 distinct phosphotyrosine (pTyr) recognition modules—the pTyr binding (PTB) domain and the Src homology 2 (SH2) domain—and multiple potential sites for tyrosine (Tyr) phosphorylation (Tyr residues 239, 240, and 317). On stimulation of hematopoietic cells with interleukin 3 (IL-3), Shc becomes phosphorylated and may therefore contribute to IL-3 signaling. We investigated the interactions mediated by the Shc modular domains and pTyr sites in IL-3–dependent IC2 premast cells. The Shc PTB domain, rather than the SH2 domain, associated both in vitro and in vivo with the Tyr-phosphorylated β subunit of the IL-3 receptor and with the SH2-containing 5′ inositol phosphatase (SHIP), and it recognized specific NXXpY phosphopeptides from these binding partners. In IL-3–stimulated mast cells, Shc phosphorylation occurred primarily on Tyr239 and 317 and was dependent on a functional PTB domain. Phosphorylated Tyr317, and to a lesser extent, Tyr239, bound the Grb2 adaptor and SHIP. Furthermore, a pTyr317 Shc phosphopeptide selectively recognized Grb2, Sos1, SHIP, and the p85 subunit of phosphatidylinositol 3′ kinase from mast cells, as characterized by mass spectrometry. These results indicate that Shc undergoes an interdependent series of pTyr-mediated interactions in IL-3–stimulated mast cells, resulting in the recruitment of proteins that regulate the Ras pathway and phospholipid metabolism.

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Indirect recruitment of the signalling adaptor Shc to the fibroblast growth factor receptor 2 (FGFR2)

Biochemical Journal ◽

10.1042/bj20080887 ◽

2008 ◽

Vol 416 (2) ◽

pp. 189-199 ◽

Cited By ~ 10

Author(s):

Annika C. Schüller ◽

Zamal Ahmed ◽

James A. Levitt ◽

Kin M. Suen ◽

Klaus Suhling ◽

...

Keyword(s):

Growth Factor ◽

Fibroblast Growth Factor Receptor ◽

Growth Factor Receptor ◽

Adaptor Protein ◽

Sh2 Domain ◽

Signalling Pathways ◽

Fibroblast Growth ◽

Src Homology ◽

Receptor Family ◽

Ptb Domain

The adaptor protein Shc (Src homology and collagen-containing protein) plays an important role in the activation of signalling pathways downstream of RTKs (receptor tyrosine kinases) regulating diverse cellular functions, such as differentiation, adhesion, migration and mitogenesis. Despite being phosphorylated downstream of members of the FGFR (fibroblast growth factor receptor) family, a direct interaction of Shc with this receptor family has not been described to date. Various studies have suggested potential binding sites for the Shc PTB domain (phosphotyrosine-binding domain) and/or the SH2 (Src homology 2) domain on FGFR1, but no interaction of full-length Shc with these sites has been reported in vivo. In the present study, we investigated the importance of the SH2 domain and the PTB domain in recruitment of Shc to FGFR2(IIIc) to characterize the interaction of these two proteins. Confocal microscopy revealed extensive co-localization of Shc with FGFR2. The PTB domain was identified as the critical component of Shc which mediates membrane localization. Results from FLIM (fluorescence lifetime imaging microscopy) revealed that the interaction between Shc and FGFR2 is indirect, suggesting that the adaptor protein forms part of a signalling complex containing the receptor. We identified the non-RTK Src as a protein which potentially mediates the formation of such a ternary complex. Although an interaction between Src and Shc has been described previously, in the present study we implicate the Shc SH2 domain as a novel mediator of this association. The recruitment of Shc to FGFR2 via an indirect mechanism provides new insight into the regulation of protein assembly and activation of various signalling pathways downstream of this RTK.

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