p56lck, LFA-1 and PI3K but not SHP-2 interact with GM1- or GM3-enriched microdomains in a CD4–p56lck association-dependent manner

We previously showed that the association of CD4 and GM3 ganglioside induced by CD4 ligand binding was required for the down-regulation of adhesion and that aggregation of ganglioside-enriched domains was accompanied by transient co-localization of LFA-1 (lymphocyte function-associated antigen-1), PI3K (phosphoinositide 3-kinase) and CD4. We also showed that these proteins co-localized with the GM1 ganglioside that partially co-localized with GM3 in these domains. In the present study, we show that CD4–p56lck association in CD4 signalling is required for the redistribution of p56lck, PI3K and LFA-1 in ganglioside-enriched domains, since ganglioside aggregation and recruitment of these proteins were not observed in a T-cell line (A201) expressing the mutant form of CD4 that does not bind p56lck. In addition, we show that although these proteins associated in different ways with GM1 and GM3, all of the associations were dependent on CD4–p56lck association. Gangliosides could associate with these proteins that differ in affinity binding and could be modified following CD4 signalling. Our results suggest that through these associations, gangliosides transiently sequestrate these proteins and consequently inhibit LFA-1-dependent adhesion. Furthermore, while structural diversity of gangliosides may allow association with distinct proteins, we show that the tyrosine phosphatase SHP-2 (Src homology 2 domain-containing protein tyrosine phosphatase 2), also required for the down-regulation of LFA-1-dependent adhesion, transiently and partially co-localized with PI3K and p56lck in detergent-insoluble membranes without association with GM1 or GM3. We propose that CD4 ligation and binding with p56lck and their interaction with GM3 and/or GM1 gangliosides induce recruitment of distinct proteins important for CD4 signalling to form a multimolecular signalling complex.

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Role of SHPS-1 in the Regulation of Insulin-like Growth Factor I–stimulated Shc and Mitogen-activated Protein Kinase Activation in Vascular Smooth Muscle Cells

Molecular Biology of the Cell ◽

10.1091/mbc.e04-10-0918 ◽

2005 ◽

Vol 16 (7) ◽

pp. 3353-3364 ◽

Cited By ~ 48

Author(s):

Yan Ling ◽

Laura A. Maile ◽

Jaroslava Lieskovska ◽

Jane Badley-Clarke ◽

David R. Clemmons

Keyword(s):

Tyrosine Phosphatase ◽

Mitogen Activated Protein Kinase ◽

Mutant Form ◽

Mapk Phosphorylation ◽

Src Homology 2 ◽

Igf I ◽

Mitogen Activated Protein ◽

Src Homology ◽

Src Homology 2 Domain

Insulin-like growth factor I (IGF-I) stimulates smooth muscle cell (SMC) proliferation, and the mitogen-activated protein kinase (MAPK) pathway plays an important role in mediating IGF-I–induced mitogenic signaling. Our prior studies have shown that recruitment of Src homology 2 domain tyrosine phosphatase (SHP-2) to the membrane scaffolding protein Src homology 2 domain–containing protein tyrosine phosphatase substrate-1 (SHPS-1) is required for IGF-I–dependent MAPK activation. The current studies were undertaken to define the upstream signaling components that are required for IGF-I–stimulated MAPK activation and the role of SHPS-1 in regulating this process. The results show that IGF-I–induced Shc phosphorylation and its subsequent binding to Grb2 is required for sustained phosphorylation of MAPK and increased cell proliferation in SMCs. Furthermore, for Shc to be phosphorylated in response to IGF-I requires that Shc must associate with SHPS-1 and this association is mediated in part by SHP-2. Preincubation of cells with a peptide that contains a phospho-tyrosine binding motif sequence derived from SHPS-1 inhibited IGF-I–stimulated SHP-2 transfer to SHPS-1, the association of Shc with SHPS-1, and IGF-I–dependent Shc phosphorylation. Expression of an SHPS-1 mutant that did not bind to Shc or SHP-2 resulted in decreased Shc and MAPK phosphorylation in response to IGF-I. In addition, SMCs expressing a mutant form of the β3 subunit of the αVβ3, which results in impairment of SHP-2 transfer to SHPS-1, also showed attenuated IGF-I–dependent Shc and MAPK phosphorylation. Further analysis showed that Shc and SHP-2 can be coimmunoprecipitated after IGF-I stimulation. A cell-permeable peptide that contained a polyproline sequence from Shc selectively inhibited Shc/SHP-2 association and impaired Shc but not SHP-2 binding to SHPS-1. Exposure to this peptide also inhibited IGF-I–stimulated Shc and MAPK phosphorylation. Cells expressing a mutant form of Shc with the four prolines substituted with alanines showed no Shc/SHPS-1 association in response to IGF-I. We conclude that SHPS-1 functions as an anchor protein that recruits both Shc and SHP-2 and that their recruitment is necessary for IGF-I–dependent Shc phosphorylation, which is required for an optimal mitogenic response in SMCs.

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Investigating the reason for loss-of-function of Src homology 2 domain-containing protein tyrosine phosphatase 2 (SHP2) caused by Y279C mutation through molecular dynamics simulation

Journal of Biomolecular Structure and Dynamics ◽

10.1080/07391102.2019.1634641 ◽

2019 ◽

Vol 38 (9) ◽

pp. 2509-2520 ◽

Cited By ~ 2

Author(s):

Wen-Shan Liu ◽

Rui-Rui Wang ◽

Wei-Ya Li ◽

Mei Rong ◽

Chi-Lu Liu ◽

...

Keyword(s):

Molecular Dynamics ◽

Molecular Dynamics Simulation ◽

Protein Tyrosine Phosphatase ◽

Tyrosine Phosphatase ◽

Dynamics Simulation ◽

Loss Of Function ◽

Protein Tyrosine ◽

Src Homology 2 ◽

Src Homology ◽

Src Homology 2 Domain

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Synthesis and biological evaluation of 2,5-diaryl-1,3,4-oxadiazole derivatives as novel Src homology 2 domain-containing protein tyrosine phosphatase 2 (SHP2) inhibitors

Bioorganic Chemistry ◽

10.1016/j.bioorg.2021.105384 ◽

2021 ◽

pp. 105384

Author(s):

Xiang-Dong Meng ◽

Li-Xin Gao ◽

Zhi-Jia Wang ◽

Bo Feng ◽

Chun Zhang ◽

...

Keyword(s):

Protein Tyrosine Phosphatase ◽

Tyrosine Phosphatase ◽

Biological Evaluation ◽

Protein Tyrosine ◽

Src Homology 2 ◽

Src Homology ◽

Oxadiazole Derivatives ◽

Src Homology 2 Domain ◽

Synthesis And Biological Evaluation

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Inhibition of Src homology 2 domain containing protein tyrosine phosphatase as the possible mechanism of metformin-assisted amelioration of obesity induced insulin resistance in high fat diet fed C57BL/6J mice

Biochemical and Biophysical Research Communications ◽

10.1016/j.bbrc.2017.04.012 ◽

2017 ◽

Vol 487 (1) ◽

pp. 54-61 ◽

Cited By ~ 5

Author(s):

Yadhu Sharma ◽

Samina Bashir ◽

Ansarullah ◽

Mohemmed Faraz Khan ◽

Altaf Ahmad ◽

...

Keyword(s):

Insulin Resistance ◽

Protein Tyrosine Phosphatase ◽

High Fat Diet ◽

Tyrosine Phosphatase ◽

High Fat ◽

Protein Tyrosine ◽

Src Homology 2 ◽

Src Homology ◽

Src Homology 2 Domain

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Design, synthesis, biological evaluation, common feature pharmacophore model and molecular dynamics simulation studies of ethyl 4-(phenoxymethyl)-2-phenylthiazole-5-carboxylate as Src homology-2 domain containing protein tyrosine phosphatase-2 (SHP2) inhibitors

Journal of Biomolecular Structure and Dynamics ◽

10.1080/07391102.2020.1726817 ◽

2020 ◽

pp. 1-15 ◽

Cited By ~ 3

Author(s):

Jingwei Wu ◽

Weiya Li ◽

Zhihui Zheng ◽

Xinhua Lu ◽

Huan Zhang ◽

...

Keyword(s):

Protein Tyrosine Phosphatase ◽

Tyrosine Phosphatase ◽

Pharmacophore Model ◽

Biological Evaluation ◽

Dynamics Simulation ◽

Simulation Studies ◽

Design Synthesis ◽

Src Homology 2 ◽

Src Homology ◽

Src Homology 2 Domain

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Chicken Ig-Like Receptor B2, a Member of a Multigene Family, Is Mainly Expressed on B Lymphocytes, Recruits Both Src Homology 2 Domain Containing Protein Tyrosine Phosphatase (SHP)-1 and SHP-2, and Inhibits Proliferation

The Journal of Immunology ◽

10.4049/jimmunol.173.12.7385 ◽

2004 ◽

Vol 173 (12) ◽

pp. 7385-7393 ◽

Cited By ~ 29

Author(s):

Birgit C. Viertlboeck ◽

Richard P. M. A. Crooijmans ◽

Martien A. M. Groenen ◽

Thomas W. F. Göbel

Keyword(s):

B Lymphocytes ◽

Protein Tyrosine Phosphatase ◽

Multigene Family ◽

Tyrosine Phosphatase ◽

Protein Tyrosine ◽

Src Homology 2 ◽

Src Homology ◽

Src Homology 2 Domain

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Leishmania EF-1α Activates the Src Homology 2 Domain Containing Tyrosine Phosphatase SHP-1 Leading to Macrophage Deactivation

Journal of Biological Chemistry ◽

10.1074/jbc.m209210200 ◽

2002 ◽

Vol 277 (51) ◽

pp. 50190-50197 ◽

Cited By ~ 98

Author(s):

Devki Nandan ◽

Taolin Yi ◽

Martin Lopez ◽

Crystal Lai ◽

Neil E. Reiner

Keyword(s):

Leishmania Donovani ◽

Tyrosine Phosphatase ◽

Elongation Factor ◽

Host Protein ◽

Src Homology 2 ◽

Infected Cells ◽

Src Homology ◽

Src Homology 2 Domain

The human leishmaniasis are persistent infections of macrophages caused by protozoa of the genusLeishmania.The chronic nature of these infections is in part related to induction of macrophage deactivation, linked to activation of the Src homology 2 domain containing tyrosine phosphatase-1 (SHP-1) in infected cells. To investigate the mechanism of SHP-1 activation, lysates ofLeishmania donovanipromastigotes were subjected to SHP-1 affinity chromatography and proteins bound to the matrix were sequenced by mass spectrometry. This resulted in the identification ofLeishmaniaelongation factor-1α (EF-1α) as a SHP-1-binding protein. PurifiedLeishmaniaEF-1α, but not host cell EF-1α, bound directly to SHP-1in vitroleading to its activation. Three independent lines of evidence indicated thatLeishmaniaEF-1α may be exported from the phagosome thereby enabling targeting of host SHP-1. First, cytosolic fractions prepared from macrophages infected with [35S]methionine-labeled organisms containedLeishmaniaEF-1α. Second, confocal, fluorescence microscopy usingLeishmania-specific antisera detectedLeishmaniaEF-1α in the cytosol of infected cells. Third, co-immunoprecipitation showed thatLeishmaniaEF-1α was associated with SHP-1in vivoin infected cells. Finally, introduction of purifiedLeishmaniaEF-1α, but not the corresponding host protein into macrophages activated SHP-1 and blocked the induction of inducible nitric-oxide synthase expression in response to interferon-γ. Thus,LeishmaniaEF-1α is identified as a novel SHP-1-binding and activating protein that recapitulates the deactivated phenotype of infected macrophages.

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