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Mohammed Akli Ayoub - One of the best experts on this subject based on the ideXlab platform.

  • Cell Surface Protein Protein interaction analysis with time resolved fret and snap tag technologies application to gpcr oligomerization
    Nature Methods, 2008
    Co-Authors: Damien Maurel, Laetitia Compsagrar, Carsten Brock, Marielaure Rives, Emmanuel Bourrier, Mohammed Akli Ayoub
    Abstract:

    Cell-Surface Proteins are important in Cell-Cell communication. They assemble into heterocomplexes that include different receptors and effectors. Elucidation and manipulation of such Protein complexes offers new therapeutic possibilities. We describe a methodology combining time-resolved fluorescence resonance energy transfer (FRET) with snap-tag technology to quantitatively analyze Protein-Protein interactions at the Surface of living Cells, in a high throughput-compatible format. Using this approach, we examined whether G Protein-coupled receptors (GPCRs) are monomers or assemble into dimers or larger oligomers--a matter of intense debate. We obtained evidence for the oligomeric state of both class A and class C GPCRs. We also observed different quaternary structure of GPCRs for the neurotransmitters glutamate and gamma-aminobutyric acid (GABA): whereas metabotropic glutamate receptors assembled into strict dimers, the GABA(B) receptors spontaneously formed dimers of heterodimers, offering a way to modulate G-Protein coupling efficacy. This approach will be useful in systematic analysis of Cell-Surface Protein interaction in living Cells.

  • Cell Surface Protein-Protein interaction analysis with combined time-resolved FRET and snap-tag technologies: application to GPCR oligomerization
    Nature Methods, 2008
    Co-Authors: Damien Maurel, Carsten Brock, Marielaure Rives, Emmanuel Bourrier, Mohammed Akli Ayoub, Laetitia Comps-agrar, Hervé Bazin, Norbert Tinel, Thierry Durroux, Laurent Prézeau
    Abstract:

    Cell-Surface Proteins are important in Cell-Cell communication. They assemble into heterocomplexes that include different receptors and effectors. Elucidation and manipulation of such Protein complexes offers new therapeutic possibilities. We describe a methodology combining time-resolved fluorescence resonance energy transfer (FRET) with snap-tag technology to quantitatively analyze Protein-Protein interactions at the Surface of living Cells, in a high throughput-compatible format. Using this approach, we examined whether G Protein-coupled receptors (GPCRs) are monomers or assemble into dimers or larger oligomers--a matter of intense debate. We obtained evidence for the oligomeric state of both class A and class C GPCRs. We also observed different quaternary structure of GPCRs for the neurotransmitters glutamate and gamma-aminobutyric acid (GABA): whereas metabotropic glutamate receptors assembled into strict dimers, the GABA(B) receptors spontaneously formed dimers of heterodimers, offering a way to modulate G-Protein coupling efficacy. This approach will be useful in systematic analysis of Cell-Surface Protein interaction in living Cells.

Kris Thielemans - One of the best experts on this subject based on the ideXlab platform.

  • The B-Cell Surface Protein CD72/Lyb-2 is the ligand for CD5.
    Nature, 1991
    Co-Authors: Hilde Van De Velde, Jane R. Parnes, Ilka Von Hoegen, Wei Luo, Kris Thielemans
    Abstract:

    THE glycoProtein CD5 is expressed on the Surface membrane of all mature T Cells1 and a small proportion of B lymphocytes1,2. Its exact role in immune interactions is still unknown. Studies3–10indicate that CD5 functions both in mice and humans as a receptor, delivering co-stimulatory signals to T Cells in a manner similar to CD2 (ref. 11) and CD28 (ref. 12). Anti-CDS antibodies stimulate both T-Cell proliferation mediated by CD3 in association with the T-Cell receptor and secretion of interleukin-2 and expression of its receptor, as well as inducing an increase in intraCellular Ca2+concentration (refs 5–10). To identify the ligand for CD5 we purified the human CD5 Protein, labelled it with biotin and used it as a probe. Here we report that CD5 specifically interacts with the Cell-Surface Protein CD72 exclusive to B Cells. This interaction is blocked by anti-CD72 antibodies, but not by any other anti-B-Cell antibodies. Moreover, non-B Cells (mouse L-Cell fibroblasts and human Jurkat T Cells) expressing a transfected human CD72 complementary DNA could bind to the CD5–biotin conjugate. The results demonstrate that the B-Cell Surface Protein CD72 (Lyb-2 in mice) is the ligand for CD5.

  • the b Cell Surface Protein cd72 lyb 2 is the ligand for cd5
    Nature, 1991
    Co-Authors: Hilde Van De Velde, Jane R. Parnes, Ilka Von Hoegen, Wei Luo, Kris Thielemans
    Abstract:

    THE glycoProtein CD5 is expressed on the Surface membrane of all mature T Cells1 and a small proportion of B lymphocytes1,2. Its exact role in immune interactions is still unknown. Studies3–10indicate that CD5 functions both in mice and humans as a receptor, delivering co-stimulatory signals to T Cells in a manner similar to CD2 (ref. 11) and CD28 (ref. 12). Anti-CDS antibodies stimulate both T-Cell proliferation mediated by CD3 in association with the T-Cell receptor and secretion of interleukin-2 and expression of its receptor, as well as inducing an increase in intraCellular Ca2+concentration (refs 5–10). To identify the ligand for CD5 we purified the human CD5 Protein, labelled it with biotin and used it as a probe. Here we report that CD5 specifically interacts with the Cell-Surface Protein CD72 exclusive to B Cells. This interaction is blocked by anti-CD72 antibodies, but not by any other anti-B-Cell antibodies. Moreover, non-B Cells (mouse L-Cell fibroblasts and human Jurkat T Cells) expressing a transfected human CD72 complementary DNA could bind to the CD5–biotin conjugate. The results demonstrate that the B-Cell Surface Protein CD72 (Lyb-2 in mice) is the ligand for CD5.

Jane R. Parnes - One of the best experts on this subject based on the ideXlab platform.

  • Allele-specific expression of the mouse B-Cell Surface Protein CD72 on T Cells
    Immunogenetics, 1997
    Co-Authors: William H. Robinson, Michelle M. Tutt Landolfi, Jane R. Parnes
    Abstract:

    CD72 is a 45 000 M r mouse B-Cell Surface glycoProtein involved in B-Cell proliferation and differentiation. Expression of mouse CD72 is thought to be restricted to the B-Cell lineage. We recently demonstrated that the monoclonal antibodies K10.6 and B9.689, previously defined as recognizing the mouse lymphocyte alloantigens Ly-19.2 and Ly-32.2, respectively, recognize specific alleles of CD72. Early studies using antibody-mediated cytotoxicity assays demonstrated that K10.6 and B9.689 react with B Cells, several T-Cell lines, and a subset of peripheral T Cells. These findings led us to consider the possibility that CD72 might also be expressed on a subset of T Cells. In this report we demonstrate that CD72 is constitutively expressed on a fraction of peripheral T Cells isolated from strains of mice expressing the CD72 b allele, but not the CD72 a or CD72 c alleles. Three days after activating T Cells with concanavalin A or plate-bound CD3-specific mAb, CD72 is expressed on a larger fraction of peripheral T Cells as well as a fraction of thymocytes from mouse strains expressing the CD72 b allele. CD72 is expressed on both the CD4+ and CD8+ thymocyte and peripheral T-Cell subsets. No CD72 expression is detected on activated thymocytes or peripheral T Cells from mouse strains expressing the CD72 a or CD72 c alleles. Expression of CD72 b on peripheral T Cells was confirmed by northern blot analysis demonstrating CD72 mRNA expression. These results demonstrate that CD72 expression is not restricted to B lineage Cells in mouse strains expressing the CD72 b allele; instead, a population of T lineage Cells in these mice also expresses CD72.

  • The B-Cell Surface Protein CD72/Lyb-2 is the ligand for CD5.
    Nature, 1991
    Co-Authors: Hilde Van De Velde, Jane R. Parnes, Ilka Von Hoegen, Wei Luo, Kris Thielemans
    Abstract:

    THE glycoProtein CD5 is expressed on the Surface membrane of all mature T Cells1 and a small proportion of B lymphocytes1,2. Its exact role in immune interactions is still unknown. Studies3–10indicate that CD5 functions both in mice and humans as a receptor, delivering co-stimulatory signals to T Cells in a manner similar to CD2 (ref. 11) and CD28 (ref. 12). Anti-CDS antibodies stimulate both T-Cell proliferation mediated by CD3 in association with the T-Cell receptor and secretion of interleukin-2 and expression of its receptor, as well as inducing an increase in intraCellular Ca2+concentration (refs 5–10). To identify the ligand for CD5 we purified the human CD5 Protein, labelled it with biotin and used it as a probe. Here we report that CD5 specifically interacts with the Cell-Surface Protein CD72 exclusive to B Cells. This interaction is blocked by anti-CD72 antibodies, but not by any other anti-B-Cell antibodies. Moreover, non-B Cells (mouse L-Cell fibroblasts and human Jurkat T Cells) expressing a transfected human CD72 complementary DNA could bind to the CD5–biotin conjugate. The results demonstrate that the B-Cell Surface Protein CD72 (Lyb-2 in mice) is the ligand for CD5.

  • the b Cell Surface Protein cd72 lyb 2 is the ligand for cd5
    Nature, 1991
    Co-Authors: Hilde Van De Velde, Jane R. Parnes, Ilka Von Hoegen, Wei Luo, Kris Thielemans
    Abstract:

    THE glycoProtein CD5 is expressed on the Surface membrane of all mature T Cells1 and a small proportion of B lymphocytes1,2. Its exact role in immune interactions is still unknown. Studies3–10indicate that CD5 functions both in mice and humans as a receptor, delivering co-stimulatory signals to T Cells in a manner similar to CD2 (ref. 11) and CD28 (ref. 12). Anti-CDS antibodies stimulate both T-Cell proliferation mediated by CD3 in association with the T-Cell receptor and secretion of interleukin-2 and expression of its receptor, as well as inducing an increase in intraCellular Ca2+concentration (refs 5–10). To identify the ligand for CD5 we purified the human CD5 Protein, labelled it with biotin and used it as a probe. Here we report that CD5 specifically interacts with the Cell-Surface Protein CD72 exclusive to B Cells. This interaction is blocked by anti-CD72 antibodies, but not by any other anti-B-Cell antibodies. Moreover, non-B Cells (mouse L-Cell fibroblasts and human Jurkat T Cells) expressing a transfected human CD72 complementary DNA could bind to the CD5–biotin conjugate. The results demonstrate that the B-Cell Surface Protein CD72 (Lyb-2 in mice) is the ligand for CD5.

Marielaure Rives - One of the best experts on this subject based on the ideXlab platform.

  • Cell Surface Protein Protein interaction analysis with time resolved fret and snap tag technologies application to gpcr oligomerization
    Nature Methods, 2008
    Co-Authors: Damien Maurel, Laetitia Compsagrar, Carsten Brock, Marielaure Rives, Emmanuel Bourrier, Mohammed Akli Ayoub
    Abstract:

    Cell-Surface Proteins are important in Cell-Cell communication. They assemble into heterocomplexes that include different receptors and effectors. Elucidation and manipulation of such Protein complexes offers new therapeutic possibilities. We describe a methodology combining time-resolved fluorescence resonance energy transfer (FRET) with snap-tag technology to quantitatively analyze Protein-Protein interactions at the Surface of living Cells, in a high throughput-compatible format. Using this approach, we examined whether G Protein-coupled receptors (GPCRs) are monomers or assemble into dimers or larger oligomers--a matter of intense debate. We obtained evidence for the oligomeric state of both class A and class C GPCRs. We also observed different quaternary structure of GPCRs for the neurotransmitters glutamate and gamma-aminobutyric acid (GABA): whereas metabotropic glutamate receptors assembled into strict dimers, the GABA(B) receptors spontaneously formed dimers of heterodimers, offering a way to modulate G-Protein coupling efficacy. This approach will be useful in systematic analysis of Cell-Surface Protein interaction in living Cells.

  • Cell Surface Protein-Protein interaction analysis with combined time-resolved FRET and snap-tag technologies: application to GPCR oligomerization
    Nature Methods, 2008
    Co-Authors: Damien Maurel, Carsten Brock, Marielaure Rives, Emmanuel Bourrier, Mohammed Akli Ayoub, Laetitia Comps-agrar, Hervé Bazin, Norbert Tinel, Thierry Durroux, Laurent Prézeau
    Abstract:

    Cell-Surface Proteins are important in Cell-Cell communication. They assemble into heterocomplexes that include different receptors and effectors. Elucidation and manipulation of such Protein complexes offers new therapeutic possibilities. We describe a methodology combining time-resolved fluorescence resonance energy transfer (FRET) with snap-tag technology to quantitatively analyze Protein-Protein interactions at the Surface of living Cells, in a high throughput-compatible format. Using this approach, we examined whether G Protein-coupled receptors (GPCRs) are monomers or assemble into dimers or larger oligomers--a matter of intense debate. We obtained evidence for the oligomeric state of both class A and class C GPCRs. We also observed different quaternary structure of GPCRs for the neurotransmitters glutamate and gamma-aminobutyric acid (GABA): whereas metabotropic glutamate receptors assembled into strict dimers, the GABA(B) receptors spontaneously formed dimers of heterodimers, offering a way to modulate G-Protein coupling efficacy. This approach will be useful in systematic analysis of Cell-Surface Protein interaction in living Cells.

Carsten Brock - One of the best experts on this subject based on the ideXlab platform.

  • Cell Surface Protein Protein interaction analysis with time resolved fret and snap tag technologies application to gpcr oligomerization
    Nature Methods, 2008
    Co-Authors: Damien Maurel, Laetitia Compsagrar, Carsten Brock, Marielaure Rives, Emmanuel Bourrier, Mohammed Akli Ayoub
    Abstract:

    Cell-Surface Proteins are important in Cell-Cell communication. They assemble into heterocomplexes that include different receptors and effectors. Elucidation and manipulation of such Protein complexes offers new therapeutic possibilities. We describe a methodology combining time-resolved fluorescence resonance energy transfer (FRET) with snap-tag technology to quantitatively analyze Protein-Protein interactions at the Surface of living Cells, in a high throughput-compatible format. Using this approach, we examined whether G Protein-coupled receptors (GPCRs) are monomers or assemble into dimers or larger oligomers--a matter of intense debate. We obtained evidence for the oligomeric state of both class A and class C GPCRs. We also observed different quaternary structure of GPCRs for the neurotransmitters glutamate and gamma-aminobutyric acid (GABA): whereas metabotropic glutamate receptors assembled into strict dimers, the GABA(B) receptors spontaneously formed dimers of heterodimers, offering a way to modulate G-Protein coupling efficacy. This approach will be useful in systematic analysis of Cell-Surface Protein interaction in living Cells.

  • Cell Surface Protein-Protein interaction analysis with combined time-resolved FRET and snap-tag technologies: application to GPCR oligomerization
    Nature Methods, 2008
    Co-Authors: Damien Maurel, Carsten Brock, Marielaure Rives, Emmanuel Bourrier, Mohammed Akli Ayoub, Laetitia Comps-agrar, Hervé Bazin, Norbert Tinel, Thierry Durroux, Laurent Prézeau
    Abstract:

    Cell-Surface Proteins are important in Cell-Cell communication. They assemble into heterocomplexes that include different receptors and effectors. Elucidation and manipulation of such Protein complexes offers new therapeutic possibilities. We describe a methodology combining time-resolved fluorescence resonance energy transfer (FRET) with snap-tag technology to quantitatively analyze Protein-Protein interactions at the Surface of living Cells, in a high throughput-compatible format. Using this approach, we examined whether G Protein-coupled receptors (GPCRs) are monomers or assemble into dimers or larger oligomers--a matter of intense debate. We obtained evidence for the oligomeric state of both class A and class C GPCRs. We also observed different quaternary structure of GPCRs for the neurotransmitters glutamate and gamma-aminobutyric acid (GABA): whereas metabotropic glutamate receptors assembled into strict dimers, the GABA(B) receptors spontaneously formed dimers of heterodimers, offering a way to modulate G-Protein coupling efficacy. This approach will be useful in systematic analysis of Cell-Surface Protein interaction in living Cells.