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gst tagged sh2 domains  (Santa Cruz Biotechnology)


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    Structured Review

    Santa Cruz Biotechnology gst tagged sh2 domains
    TSAd interactions with VEGFR2 and Src. (A) Immobilized peptides corresponding to tyrosine phosphorylation sites in VEGFR2 and TSAd were probed using TSAd <t>GST-SH2</t> (left) or c-Src GST-SH2 (right) domain fusion proteins. The phosphotyrosine position in VEGFR2 or TSAd is indicated to the left and the peptide sequence to the right in the figure. The figure shows representative results from three independent experiments. Line indicates the splice of two parts of the same filter aligned to one strip. (B) Immobilized peptides corresponding to proline-rich stretches in TSAd were probed with c-Src GST-SH3 domain fusion protein. Peptide sequences are shown to the right. The figure shows representative results from three independent experiments. Line indicates the splice of two parts of the same filter aligned to one strip. (C) Schematic representation of VEGFR2, TSAd, and c-Src molecular interactions.
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    Images

    1) Product Images from "VEGFR2 induces c-Src signaling and vascular permeability in vivo via the adaptor protein TSAd"

    Article Title: VEGFR2 induces c-Src signaling and vascular permeability in vivo via the adaptor protein TSAd

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20111343

    TSAd interactions with VEGFR2 and Src. (A) Immobilized peptides corresponding to tyrosine phosphorylation sites in VEGFR2 and TSAd were probed using TSAd GST-SH2 (left) or c-Src GST-SH2 (right) domain fusion proteins. The phosphotyrosine position in VEGFR2 or TSAd is indicated to the left and the peptide sequence to the right in the figure. The figure shows representative results from three independent experiments. Line indicates the splice of two parts of the same filter aligned to one strip. (B) Immobilized peptides corresponding to proline-rich stretches in TSAd were probed with c-Src GST-SH3 domain fusion protein. Peptide sequences are shown to the right. The figure shows representative results from three independent experiments. Line indicates the splice of two parts of the same filter aligned to one strip. (C) Schematic representation of VEGFR2, TSAd, and c-Src molecular interactions.
    Figure Legend Snippet: TSAd interactions with VEGFR2 and Src. (A) Immobilized peptides corresponding to tyrosine phosphorylation sites in VEGFR2 and TSAd were probed using TSAd GST-SH2 (left) or c-Src GST-SH2 (right) domain fusion proteins. The phosphotyrosine position in VEGFR2 or TSAd is indicated to the left and the peptide sequence to the right in the figure. The figure shows representative results from three independent experiments. Line indicates the splice of two parts of the same filter aligned to one strip. (B) Immobilized peptides corresponding to proline-rich stretches in TSAd were probed with c-Src GST-SH3 domain fusion protein. Peptide sequences are shown to the right. The figure shows representative results from three independent experiments. Line indicates the splice of two parts of the same filter aligned to one strip. (C) Schematic representation of VEGFR2, TSAd, and c-Src molecular interactions.

    Techniques Used: Phospho-proteomics, Sequencing, Stripping Membranes

    Related Articles

    In Vitro:

    Article Title: β-Hydroxybutyrate promotes cancer metastasis through β-hydroxybutyrylation-dependent stabilization of Snail
    Article Snippet: .. To detect the interaction of Snail and CBP in vitro, GST or Snail-GST protein was incubated with Glutathione-Sepharose beads (Santa Cruz, sc-2009) at 4 °C for 3 h. The complexes were washed with buffer 5 times and incubated with His-tagged protein, and the mixture was incubated at 4 °C for another 3 h. After washing, the pulled-down proteins were analyzed by immunoblotting with anti-GST antibody and anti-His tag antibody. ..

    Incubation:

    Article Title: β-Hydroxybutyrate promotes cancer metastasis through β-hydroxybutyrylation-dependent stabilization of Snail
    Article Snippet: .. To detect the interaction of Snail and CBP in vitro, GST or Snail-GST protein was incubated with Glutathione-Sepharose beads (Santa Cruz, sc-2009) at 4 °C for 3 h. The complexes were washed with buffer 5 times and incubated with His-tagged protein, and the mixture was incubated at 4 °C for another 3 h. After washing, the pulled-down proteins were analyzed by immunoblotting with anti-GST antibody and anti-His tag antibody. ..

    Article Title: Site-specific phosphorylation of LRP regulates FLC chromatin looping and flowering
    Article Snippet: Protein concentrations were measured using the Bradford assay (Bio-Rad). .. GST-LRP N (200 ng) was incubated with 500 μg of total protein extract in the reaction buffer [20 mM tris-HCl (pH 7.5), 10 mM MgCl 2 , 1 mM DTT, and 1 mM ATP] with or without 50 μM MG132 at 37°C for 60, 120, and 180 min. GST-LRP N protein levels in different reactions were analyzed by SDS-PAGE followed by detection using anti-GST antibody (Santa Cruz Biotechnology). .. Band intensities were quantified using ImageJ software (US National Institutes of Health).

    Western Blot:

    Article Title: β-Hydroxybutyrate promotes cancer metastasis through β-hydroxybutyrylation-dependent stabilization of Snail
    Article Snippet: .. To detect the interaction of Snail and CBP in vitro, GST or Snail-GST protein was incubated with Glutathione-Sepharose beads (Santa Cruz, sc-2009) at 4 °C for 3 h. The complexes were washed with buffer 5 times and incubated with His-tagged protein, and the mixture was incubated at 4 °C for another 3 h. After washing, the pulled-down proteins were analyzed by immunoblotting with anti-GST antibody and anti-His tag antibody. ..

    Article Title: Phosphorylation-Induced Degradation of Pepper Dehydration-Related Homeobox Gene 12 (CaDRHB12), a Negative Regulator of Drought Response in Pepper, by CaSnRK2.6 Enhances Drought Tolerance.
    Article Snippet: .. Western blotting was conducted using GST (Santa Cruz Biotechnology, Dallas, TX, USA) and MBP (New England Biolabs) antibodies. ..

    Article Title: Cytomegalovirus-encoded immediate early 1 protein perturbs neural progenitor proliferation via interfering with host PML–DISC1 interaction
    Article Snippet: .. Bound recombinant proteins were detected with anti-GST antisera (Santa Cruz Biotechnologies) and a complementary horseradish peroxidase-coupled secondary antibody for immunoblotting. .. Cells or tissues were lysed in lysis buffer (150 mM NaCl, 50 mM Tris–HCl, pH 7.5, 1% Triton X-100) containing a protease inhibitor mixture (Roche Applied Sciences).

    Article Title: Phosphorylation‐Induced Degradation of Pepper Dehydration‐Related Homeobox Gene 12 (CaDRHB12), a Negative Regulator of Drought Response in Pepper, by CaSnRK2.6 Enhances Drought Tolerance
    Article Snippet: .. Western blotting was conducted using GST (Santa Cruz Biotechnology, Dallas, TX, USA) and MBP (New England Biolabs) antibodies. ..

    Article Title: DISC1-PML protein interaction for congenital CMV infection-induced cortical neural progenitor deficit: perturbance of host signaling via viral IE1
    Article Snippet: .. Bound recombinant proteins were detected with anti-GST antisera (Santa Cruz Biotechnologies) and a complementary HRP-coupled secondary antibody for immunoblotting. .. Cells or tissues were lysed in lysis buffer (150 mM NaCl, 50 mM Tris-HCl, pH 7.5, 1% Triton X-100) containing a protease inhibitor mixture (Roche Applied Sciences).

    Chromatin Immunoprecipitation:

    Article Title: Procollagen IIA mediates positive feedback control of the mouse cardiogenic transcriptional network.
    Article Snippet: .. ChIP was performed with anti- NKX2- 5 (sc- 8697, sc- 4033) and anti- GATA4 (sc- 25310, Santa Cruz Biotechnology) antibodies. ..

    Recombinant:

    Article Title: Cytomegalovirus-encoded immediate early 1 protein perturbs neural progenitor proliferation via interfering with host PML–DISC1 interaction
    Article Snippet: .. Bound recombinant proteins were detected with anti-GST antisera (Santa Cruz Biotechnologies) and a complementary horseradish peroxidase-coupled secondary antibody for immunoblotting. .. Cells or tissues were lysed in lysis buffer (150 mM NaCl, 50 mM Tris–HCl, pH 7.5, 1% Triton X-100) containing a protease inhibitor mixture (Roche Applied Sciences).

    Article Title: DISC1-PML protein interaction for congenital CMV infection-induced cortical neural progenitor deficit: perturbance of host signaling via viral IE1
    Article Snippet: .. Bound recombinant proteins were detected with anti-GST antisera (Santa Cruz Biotechnologies) and a complementary HRP-coupled secondary antibody for immunoblotting. .. Cells or tissues were lysed in lysis buffer (150 mM NaCl, 50 mM Tris-HCl, pH 7.5, 1% Triton X-100) containing a protease inhibitor mixture (Roche Applied Sciences).

    SDS Page:

    Article Title: Site-specific phosphorylation of LRP regulates FLC chromatin looping and flowering
    Article Snippet: Protein concentrations were measured using the Bradford assay (Bio-Rad). .. GST-LRP N (200 ng) was incubated with 500 μg of total protein extract in the reaction buffer [20 mM tris-HCl (pH 7.5), 10 mM MgCl 2 , 1 mM DTT, and 1 mM ATP] with or without 50 μM MG132 at 37°C for 60, 120, and 180 min. GST-LRP N protein levels in different reactions were analyzed by SDS-PAGE followed by detection using anti-GST antibody (Santa Cruz Biotechnology). .. Band intensities were quantified using ImageJ software (US National Institutes of Health).

    other:

    Article Title: S100A14: A novel negative regulator of cancer stemness and immune evasion by inhibiting STAT3-mediated programmed death-ligand 1 expression n colorectal cancer
    Article Snippet: Target Vendor Catalogue Number Clone Dilution ratio Application STAT3 Santa Cruz sc-482 C-20 1:1,000 (WB) 1:200 (IP) WB, IF, IP STAT3 Cell signaling 9139 124H6 1:200 (IF) 1: 500 (IHC) IF, IHC S100A14 Proteintech 10489-1-AP 1:1,000 (WB) 1:200 (IF) 1:200 (IHC) WB, IF, IHC pSTAT3 Cell signaling 9145 D3A7 1:1,000 WB Ac-STAT3 Cell signaling 2523 1:1,000 WB Oct4 Abcam ab-19857 1:1,000 WB Nanog Cell signaling 4903 D73G4 1:1,000 WB Sox2 Abcam 97959 1:1,000 WB cMyc Santa Cruz sc-40 9E10 1:1,000 WB Cleaved caspase-3 Cell signaling 9661 1:1,000 (WB) 1:200 (IHC) WB, IHC Cleaved PARP Cell signaling 9544 1:1,000 WB PD-L1 Cell signaling 13684 E1L3N 1:1,000 WB PD-L2 Cell signaling 82723 D7U8C 1:1,000 WB His Santa Cruz sc-8036 H-3 1:1,000 WB GST Santa Cruz sc-459 Z-5 1:1,000 WB Purified antihuman CD274 Biolegend 329702 29E.2A3 1:100 / 1.5×106 cells (FACS, FC) 1:50 (IF, IHC) 0.5 μg (Blockade) FACS, IF, IHC, Blockade PE anti-mouse CD274 Biolegend 124307 10F.9G2 1:100 / 1.5×106 cells FACS Purified antimouse CD274 Biolegend 124302 10F.9G2 1:50 IF - 3 - Anti-mouse PDL1 InVivoMAb BE0101 10F.9G2 8 μg/mL (WB) 10 μg/mL (blockade) WB, Blockade FITC antihuman CD44 BD Pharmingen 555478 G44-2 1:100/ test (~ 107 cells) FACS PE anti-human CD133 Miltenyl Biotec 130-090-853 293C3 1:100/ test (~ 107 cells) FACS FITC antimouse CD44 Biolegend 103005 IM7 1:100 / 1.5×106 cells FACS PE anti-mouse CD133 Biolegend 141203 315-2C11 1:100 / 1.5×106 cells FACS PE anti-mouse IFN-γ Biolegend 505807 XMG1.2 1:100 / 1.5×106 cells FC APC anti-mouse CD8a BioGems 10112-80- 100 2.43 1:200 / 1.5 ×106 cells FC PerCP/Cy5.5 anti-mouse CD3e BioGems 05122-70-100 145-2C11 1:200 / 1.5 ×106 cells FC Granzyme B R&D AF-1865 1:200 IHC Ubiquitin Santa Cruz sc-8017 P4D1 1:1,000 WB Purified Mouse IgG2b, κ Isotype Ctrl Antibody Biolegend 402201 27-35 1:100 / 1.5 ×10 6 cells FC HRP-conjugated goat anti-mouse igG GeneTex 213111-01 1:5,000 WB HRP-conjugated goat anti-rabbit igG GeneTex 213110-01 1:5,000 WB Alexa Fluor 594- conjugated donkey antirabbit igG Thermo Fisher Scientific A21207 1:1,000 IF Alexa Fluor 633- conjugated goat anti-mouse igG Thermo Fisher Scientific A21052 1:200 FACS - 4 - Alexa Fluor 488- conjugated goat anti-mouse igG Thermo Fisher Scientific A11001 1:1,000 FACS Horse Anti-Goat IgG (H+L), Biotinylated Vector lab BA-9500 1:1,000 IHC Goat Anti-Rabbit IgG (H+L), Biotinylated Vector lab BA-1000 1:1,000 IHC Horse antiMouse IgG (H+L) Biotinylated Vector lab BA-2000 1:1,000 IHC 1)Application - WB: Western blot analysis; IF: Immunofluorescence staining; IHC: Immunohistochemistry; IP: Immunoprecipitation; FACS: Fluorescence-Activated Cell Sorting, FC: Flow cytometry - 5 - Table S2.



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    Image Search Results


    Src SH2 binding to cortactin does not involve tyrosine phosphorylation and binds cortactin repeats 1 and 5. (A) GST and GST-Src SH2 affinity precipitation from MTLn3 cells evaluated for cortactin tyrosine phosphorylation. The ratio of phosphorylated cortactin levels and the normalized amounts of total precipitated cortactin are indicated. (B) Src SH2 far western analysis of FLAG-tagged recombinant wild-type and cortactin phosphorylation mutants. TYM, triple tyrosine mutant. (C) Affinity precipitation analysis of FLAG-tagged recombinant wild-type and cortactin phosphorylation mutants from extracts with GST-Src SH2 domain. (D) Affinity precipitation of non-phosphorylated, recombinant cortactin with GST and GST-Src SH2. Normalized intensity levels are shown relative to GST control. (E) GST-SH2 domain far western blotting of the cortactin NTA and repeats region. CT, C terminus; LC, IgG light chain; NT, N terminus. Arrows denote position of IgG heavy chain (HC) recognized by cross reactivity with secondary antibodies during the blotting process. Asterisks indicate the positions of recombinant cortactin proteins. (F) GST-SH2 far western analysis of cortactin deletion cortactin constructs. (G) Far western binding of the GST-Src SH2 domain to tandem cortactin repeat chimeric constructs.

    Journal: Journal of Cell Science

    Article Title: Src binds cortactin through an SH2 domain cystine-mediated linkage

    doi: 10.1242/jcs.121046

    Figure Lengend Snippet: Src SH2 binding to cortactin does not involve tyrosine phosphorylation and binds cortactin repeats 1 and 5. (A) GST and GST-Src SH2 affinity precipitation from MTLn3 cells evaluated for cortactin tyrosine phosphorylation. The ratio of phosphorylated cortactin levels and the normalized amounts of total precipitated cortactin are indicated. (B) Src SH2 far western analysis of FLAG-tagged recombinant wild-type and cortactin phosphorylation mutants. TYM, triple tyrosine mutant. (C) Affinity precipitation analysis of FLAG-tagged recombinant wild-type and cortactin phosphorylation mutants from extracts with GST-Src SH2 domain. (D) Affinity precipitation of non-phosphorylated, recombinant cortactin with GST and GST-Src SH2. Normalized intensity levels are shown relative to GST control. (E) GST-SH2 domain far western blotting of the cortactin NTA and repeats region. CT, C terminus; LC, IgG light chain; NT, N terminus. Arrows denote position of IgG heavy chain (HC) recognized by cross reactivity with secondary antibodies during the blotting process. Asterisks indicate the positions of recombinant cortactin proteins. (F) GST-SH2 far western analysis of cortactin deletion cortactin constructs. (G) Far western binding of the GST-Src SH2 domain to tandem cortactin repeat chimeric constructs.

    Article Snippet: Screening of TransignalTM SH2 Domain Arrays (Panomics Cat. NO. MA3040) was conducted using 1.0 mg of each cortactin peptide according to the manufacturer's protocol.

    Techniques: Binding Assay, Phospho-proteomics, Affinity Precipitation, Western Blot, Recombinant, Mutagenesis, Control, Far Western Blot, Construct

    Cortactin cysteines 112 and 246 are required for Src SH2 domain binding. (A) Alignment of cortactin repeats denoting C112 and C246. (B,C) Far western blotting of GST-Src SH2 domain with cortactin cysteine to alanine mutants. CT, C terminus; HC, IgG heavy chain; LC, IgG light chain; NT, N terminus; WT; full-length wild-type cortactin. Dashed line separates HC from chimeric cortactin proteins (asterisks) due to similar molecular weights. (D) Far western analysis of Src SH2 domain binding to the C112/C246A cortactin double cysteine mutant (DCM). (E) Affinity precipitation analysis of Src SH2 domain binding to the C112/C246A cortactin double cysteine mutant (DCM).

    Journal: Journal of Cell Science

    Article Title: Src binds cortactin through an SH2 domain cystine-mediated linkage

    doi: 10.1242/jcs.121046

    Figure Lengend Snippet: Cortactin cysteines 112 and 246 are required for Src SH2 domain binding. (A) Alignment of cortactin repeats denoting C112 and C246. (B,C) Far western blotting of GST-Src SH2 domain with cortactin cysteine to alanine mutants. CT, C terminus; HC, IgG heavy chain; LC, IgG light chain; NT, N terminus; WT; full-length wild-type cortactin. Dashed line separates HC from chimeric cortactin proteins (asterisks) due to similar molecular weights. (D) Far western analysis of Src SH2 domain binding to the C112/C246A cortactin double cysteine mutant (DCM). (E) Affinity precipitation analysis of Src SH2 domain binding to the C112/C246A cortactin double cysteine mutant (DCM).

    Article Snippet: Screening of TransignalTM SH2 Domain Arrays (Panomics Cat. NO. MA3040) was conducted using 1.0 mg of each cortactin peptide according to the manufacturer's protocol.

    Techniques: Binding Assay, Far Western Blot, Western Blot, Mutagenesis, Affinity Precipitation

    Cysteine-containing cortactin peptides dock within the Src SH2 phosphotyrosine binding region. (A) Molecular modelling of the Src SH2 domain with phosphorylated Src and cortactin pentapeptides. Enlarged views show position of Src R175, Src C185 and the respective central Src or cortactin peptide residues. (B) Calculated binding energies for each peptide docking condition shown in (A).

    Journal: Journal of Cell Science

    Article Title: Src binds cortactin through an SH2 domain cystine-mediated linkage

    doi: 10.1242/jcs.121046

    Figure Lengend Snippet: Cysteine-containing cortactin peptides dock within the Src SH2 phosphotyrosine binding region. (A) Molecular modelling of the Src SH2 domain with phosphorylated Src and cortactin pentapeptides. Enlarged views show position of Src R175, Src C185 and the respective central Src or cortactin peptide residues. (B) Calculated binding energies for each peptide docking condition shown in (A).

    Article Snippet: Screening of TransignalTM SH2 Domain Arrays (Panomics Cat. NO. MA3040) was conducted using 1.0 mg of each cortactin peptide according to the manufacturer's protocol.

    Techniques: Binding Assay

    Binding and phosphorylation of cortactin by Src is redox dependent and requires Src C185. (A) Co-immunoprecipitation (IP) of cortactin with Src followed by analysis under reducing (R) and non-reducing (NR) conditions. Asterisks denote equivalent bands in cortactin and Src immunoblots. (B) Co-immunoprecipitation of Src with cortactin followed by analysis under reducing (R) and non-reducing (NR) conditions. Asterisks denote equivalent bands in cortactin and Src immunoblots. (C) Phosphorylation of cortactin by Src in the absence and presence of DTT. (D) Far western analysis of GST-Src SH2 and C185A. (E) Affinity precipitation analysis of FAK and cortactin binding to GST-Src SH2 C185A.

    Journal: Journal of Cell Science

    Article Title: Src binds cortactin through an SH2 domain cystine-mediated linkage

    doi: 10.1242/jcs.121046

    Figure Lengend Snippet: Binding and phosphorylation of cortactin by Src is redox dependent and requires Src C185. (A) Co-immunoprecipitation (IP) of cortactin with Src followed by analysis under reducing (R) and non-reducing (NR) conditions. Asterisks denote equivalent bands in cortactin and Src immunoblots. (B) Co-immunoprecipitation of Src with cortactin followed by analysis under reducing (R) and non-reducing (NR) conditions. Asterisks denote equivalent bands in cortactin and Src immunoblots. (C) Phosphorylation of cortactin by Src in the absence and presence of DTT. (D) Far western analysis of GST-Src SH2 and C185A. (E) Affinity precipitation analysis of FAK and cortactin binding to GST-Src SH2 C185A.

    Article Snippet: Screening of TransignalTM SH2 Domain Arrays (Panomics Cat. NO. MA3040) was conducted using 1.0 mg of each cortactin peptide according to the manufacturer's protocol.

    Techniques: Binding Assay, Phospho-proteomics, Immunoprecipitation, Western Blot, Affinity Precipitation

    Src C185 forms a cystine bond with cortactin C112 and C246. (A) Sequence of the predicted Src SH2 domain tryptic fragment containing C185. Predicted cystine bonding between Src C185 and the cortactin C112 and C246 tryptic peptides with predicted masses are shown below. (B–D) Extracted ion chromatogram (left) and ion fragmentation spectra (right) from tandem LC-MS/MS of the GST-Src SH2 domain (B), the GST-SH2 domain with the cortactin C112 peptide (C) and the GST-SH2 domain with cortactin C246 peptide (D). Spectra were enlarged to indicate the position of the Src C185 b4 ion (boxed in red).

    Journal: Journal of Cell Science

    Article Title: Src binds cortactin through an SH2 domain cystine-mediated linkage

    doi: 10.1242/jcs.121046

    Figure Lengend Snippet: Src C185 forms a cystine bond with cortactin C112 and C246. (A) Sequence of the predicted Src SH2 domain tryptic fragment containing C185. Predicted cystine bonding between Src C185 and the cortactin C112 and C246 tryptic peptides with predicted masses are shown below. (B–D) Extracted ion chromatogram (left) and ion fragmentation spectra (right) from tandem LC-MS/MS of the GST-Src SH2 domain (B), the GST-SH2 domain with the cortactin C112 peptide (C) and the GST-SH2 domain with cortactin C246 peptide (D). Spectra were enlarged to indicate the position of the Src C185 b4 ion (boxed in red).

    Article Snippet: Screening of TransignalTM SH2 Domain Arrays (Panomics Cat. NO. MA3040) was conducted using 1.0 mg of each cortactin peptide according to the manufacturer's protocol.

    Techniques: Sequencing, Liquid Chromatography with Mass Spectroscopy

    Observed and predicted ion masses of  GST-Src-SH2,  GST-Src-SH2 + cortactin C112 and GST-Src-SH2 + cortactin C246 tryptic peptides

    Journal: Journal of Cell Science

    Article Title: Src binds cortactin through an SH2 domain cystine-mediated linkage

    doi: 10.1242/jcs.121046

    Figure Lengend Snippet: Observed and predicted ion masses of GST-Src-SH2, GST-Src-SH2 + cortactin C112 and GST-Src-SH2 + cortactin C246 tryptic peptides

    Article Snippet: Screening of TransignalTM SH2 Domain Arrays (Panomics Cat. NO. MA3040) was conducted using 1.0 mg of each cortactin peptide according to the manufacturer's protocol.

    Techniques:

    Model of cysteine-mediated interactions in cortactin regulation. (A) Model of cysteine-based cortactin activation and phosphorylation by Src. (B) Phylogenetic co-conservation of cortactin C112/246 and Src C185. Conserved cysteines are in red and the homologous positions highlighted in yellow. (C) Cartoon representation of Src SH2 binding to phosphotyrosine and cysteine residues. Src amino acids 172–191 within the SH2 domain binding pocket are in white; interacting arginine 175 and cysteine 185 residues are in red. Cystine bonding is indicated as a red line. Phosphotyrosine and cystine binding ligands are listed. (D) Alignment of cysteine-containing SH2 domains. Domains known to bind ligands in a phosphotyrosine-independent manner are in bold italics. Cysteine residues are in red. Shading: green; hydrophobic, blue; positively charged, red; negatively charged, yellow; polar.

    Journal: Journal of Cell Science

    Article Title: Src binds cortactin through an SH2 domain cystine-mediated linkage

    doi: 10.1242/jcs.121046

    Figure Lengend Snippet: Model of cysteine-mediated interactions in cortactin regulation. (A) Model of cysteine-based cortactin activation and phosphorylation by Src. (B) Phylogenetic co-conservation of cortactin C112/246 and Src C185. Conserved cysteines are in red and the homologous positions highlighted in yellow. (C) Cartoon representation of Src SH2 binding to phosphotyrosine and cysteine residues. Src amino acids 172–191 within the SH2 domain binding pocket are in white; interacting arginine 175 and cysteine 185 residues are in red. Cystine bonding is indicated as a red line. Phosphotyrosine and cystine binding ligands are listed. (D) Alignment of cysteine-containing SH2 domains. Domains known to bind ligands in a phosphotyrosine-independent manner are in bold italics. Cysteine residues are in red. Shading: green; hydrophobic, blue; positively charged, red; negatively charged, yellow; polar.

    Article Snippet: Screening of TransignalTM SH2 Domain Arrays (Panomics Cat. NO. MA3040) was conducted using 1.0 mg of each cortactin peptide according to the manufacturer's protocol.

    Techniques: Activation Assay, Phospho-proteomics, Binding Assay