The Experts below are selected from a list of 294 Experts worldwide ranked by ideXlab platform
Carsten Watzl - One of the best experts on this subject based on the ideXlab platform.
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Current Protocols in Protein Science - Production and use of trimeric isoleucine zipper fusion proteins to study Surface Receptor ligand interactions.
Current Protocols in Protein Science, 2006Co-Authors: Carsten WatzlAbstract:Soluble recombinant forms of Surface Receptors are commonly used to study the interaction between cell Surface Receptors and their ligands. By fusing the extracellular domain of the Receptor to an isoleucine zipper (ILZ) sequence, it is possible to generate recombinant Receptor fusion proteins that form trimers in solution. These ILZ fusion proteins demonstrate several advantages over the commonly used Fc fusion proteins. This unit describes the production and purification of recombinant ILZ fusion proteins. A protocol is also provided for the use of ILZ fusion proteins to study cell Surface Receptor-ligand interactions on intact cells by flow cytometry. Keywords: Surface Receptor; soluble Receptor fusion protein; recombinant protein; Receptor-ligand interaction; trimer
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Production and use of trimeric isoleucine zipper fusion proteins to study Surface Receptor ligand interactions.
Current protocols in protein science, 2006Co-Authors: Carsten WatzlAbstract:Soluble recombinant forms of Surface Receptors are commonly used to study the interaction between cell Surface Receptors and their ligands. By fusing the extracellular domain of the Receptor to an isoleucine zipper (ILZ) sequence, it is possible to generate recombinant Receptor fusion proteins that form trimers in solution. These ILZ fusion proteins demonstrate several advantages over the commonly used Fc fusion proteins. This unit describes the production and purification of recombinant ILZ fusion proteins. A protocol is also provided for the use of ILZ fusion proteins to study cell Surface Receptor-ligand interactions on intact cells by flow cytometry.
Harold P Erickson - One of the best experts on this subject based on the ideXlab platform.
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mitogenesis cell migration and loss of focal adhesions induced by tenascin c interacting with its cell Surface Receptor annexin ii
Molecular Biology of the Cell, 1996Co-Authors: Chang Y Chung, Joanne E Murphyullrich, Harold P EricksonAbstract:Abstract In a previous study we demonstrated that the alternatively spliced region of tenascin-C, TNfnA-D, bound with high affinity to a cell Surface Receptor, annexin II. In the present study we demonstrate three changes in cellular activity that are produced by adding intact tenascin-C or TNfnA-D to cells, and we show that all three activities are blocked by antibodies against annexin II. 1) TNfnA-D added to confluent endothelial cells induced loss of focal adhesions. 2) TNfnA-D produced a mitogenic response of confluent, growth-arrested endothelial cells in 1% serum. TNfnA-D stimulated mitogenesis only when it was added to cells before or during exposure to other mitogens, such as basic fibroblast growth factor or serum. Thus the effect of TNfnA-D seems to be to facilitate the subsequent response to growth factors. 3) TNfnA-D enhanced cell migration in a cell culture wound assay. Antibodies to annexin II blocked all three cellular responses to TNfnA-D. These data show that annexin II Receptors on endothelial cells mediate several cell regulatory functions attributed to tenascin-C, potentially through modulation of intracellular signalling pathways.
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mitogenesis cell migration and loss of focal adhesions induced by tenascin c interacting with its cell Surface Receptor annexin ii
Molecular Biology of the Cell, 1996Co-Authors: Chang Y Chung, Joanne E Murphyullrich, Harold P EricksonAbstract:In a previous study we demonstrated that the alternatively spliced region of tenascin-C, TNfnA-D, bound with high affinity to a cell Surface Receptor, annexin II. In the present study we demonstrat...
William S Hlavacek - One of the best experts on this subject based on the ideXlab platform.
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modeling multivalent ligand Receptor interactions with steric constraints on configurations of cell Surface Receptor aggregates
Biophysical Journal, 2010Co-Authors: Michael I Monine, Richard G Posner, Paul B Savage, James R Faeder, William S HlavacekAbstract:We use flow cytometry to characterize equilibrium binding of a fluorophore-labeled trivalent model antigen to bivalent IgE-FceRI complexes on RBL cells. We find that flow cytometric measurements are consistent with an equilibrium model for ligand-Receptor binding in which binding sites are assumed to be equivalent and ligand-induced Receptor aggregates are assumed to be acyclic. However, this model predicts extensive Receptor aggregation at antigen concentrations that yield strong cellular secretory responses, which is inconsistent with the expectation that large Receptor aggregates should inhibit such responses. To investigate possible explanations for this discrepancy, we evaluate four rule-based models for interaction of a trivalent ligand with a bivalent cell-Surface Receptor that relax simplifying assumptions of the equilibrium model. These models are simulated using a rule-based kinetic Monte Carlo approach to investigate the kinetics of ligand-induced Receptor aggregation and to study how the kinetics and equilibria of ligand-Receptor interaction are affected by steric constraints on Receptor aggregate configurations and by the formation of cyclic Receptor aggregates. The results suggest that formation of linear chains of cyclic Receptor dimers may be important for generating secretory signals. Steric effects that limit Receptor aggregation and transient formation of small Receptor aggregates may also be important.
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modeling multivalent ligand Receptor interactions with steric constraints on configurations of cell Surface Receptor aggregates
PLOS Computational Biology, 2008Co-Authors: Michael I Monine, Richard G Posner, Paul B Savage, James R Faeder, William S HlavacekAbstract:Signal transduction generally involves multivalent protein-protein interactions, which can produce various protein complexes and post-translational modifications. The reaction networks that characterize these interactions tend to be so large as to challenge conventional simulation procedures. To address this challenge, a kinetic Monte Carlo (KMC) method has been developed that can take advantage of a model specification in terms of reaction rules for molecular interactions. A set of rules implicitly defines the reactions that can occur as a result of the interactions represented by the rules. With the rule-based KMC method, explicit generation of the underlying chemical reaction network implied by rules is avoided. Here, we apply and extend this method to characterize the interactions of a trivalent ligand with a bivalent cell-Surface Receptor. This system is also studied experimentally. We consider the following kinetic models: an equivalent-site model, an extension of this model, which takes into account steric constraints on the configurations of Receptor aggregates, and finally, a model that accounts for cyclic Receptor aggregates. Simulation results for the equivalent-site model are consistent with an equilibrium continuum model. Using these models, we investigate the effects of steric constraints and the formation of cyclic aggregates on the kinetics and equilibria of small more » and large aggregate formation and the percolation phase transition that occurs in this system. « less
Chang Y Chung - One of the best experts on this subject based on the ideXlab platform.
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mitogenesis cell migration and loss of focal adhesions induced by tenascin c interacting with its cell Surface Receptor annexin ii
Molecular Biology of the Cell, 1996Co-Authors: Chang Y Chung, Joanne E Murphyullrich, Harold P EricksonAbstract:Abstract In a previous study we demonstrated that the alternatively spliced region of tenascin-C, TNfnA-D, bound with high affinity to a cell Surface Receptor, annexin II. In the present study we demonstrate three changes in cellular activity that are produced by adding intact tenascin-C or TNfnA-D to cells, and we show that all three activities are blocked by antibodies against annexin II. 1) TNfnA-D added to confluent endothelial cells induced loss of focal adhesions. 2) TNfnA-D produced a mitogenic response of confluent, growth-arrested endothelial cells in 1% serum. TNfnA-D stimulated mitogenesis only when it was added to cells before or during exposure to other mitogens, such as basic fibroblast growth factor or serum. Thus the effect of TNfnA-D seems to be to facilitate the subsequent response to growth factors. 3) TNfnA-D enhanced cell migration in a cell culture wound assay. Antibodies to annexin II blocked all three cellular responses to TNfnA-D. These data show that annexin II Receptors on endothelial cells mediate several cell regulatory functions attributed to tenascin-C, potentially through modulation of intracellular signalling pathways.
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mitogenesis cell migration and loss of focal adhesions induced by tenascin c interacting with its cell Surface Receptor annexin ii
Molecular Biology of the Cell, 1996Co-Authors: Chang Y Chung, Joanne E Murphyullrich, Harold P EricksonAbstract:In a previous study we demonstrated that the alternatively spliced region of tenascin-C, TNfnA-D, bound with high affinity to a cell Surface Receptor, annexin II. In the present study we demonstrat...
Paul J Davis - One of the best experts on this subject based on the ideXlab platform.
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DEFINING THE ROLES OF THE CELL Surface Receptor FOR THYROID HORMONE
2006Co-Authors: Paul J Davis, Faith B Davis, Joel J BerghAbstract:DEFINING THE ROLES OF THE CELL Surface Receptor FOR THYROID HORMONE Paul J Davis Ordway Research Institute, Inc., Albany, NY 12208 USA Fax 518 641 6303, Stratton Veterans Affairs Medical Center, Wadsworth, Center of New York State Department of Health and Albany Medical College , , , email: pdavis@ordwayresearch.org Faith B. Davis Ordway Research Institute, Inc., Albany, NY 12208 USA Fax 518 641 6303 , , Joel Bergh Ordway Research Institute, Inc., Albany, NY 12208 USA Fax 518 641 6303, ,
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integrin alphavbeta3 contains a cell Surface Receptor site for thyroid hormone that is linked to activation of mitogen activated protein kinase and induction of angiogenesis
Endocrinology, 2005Co-Authors: Joel J Bergh, Lawrence Lansing, Seema Mohamed, Faith B Davis, Shaker A Mousa, Paul J DavisAbstract:Integrin αVβ3 is a heterodimeric plasma membrane protein whose several extracellular matrix protein ligands contain an RGD recognition sequence. This study identifies integrin αVβ3 as a cell Surface Receptor for thyroid hormone [l-T4 (T4)] and as the initiation site for T4-induced activation of intracellular signaling cascades. Integrin αVβ3 dissociably binds radiolabeled T4 with high affinity, and this binding is displaced by tetraiodothyroacetic acid, αVβ3 antibodies, and an integrin RGD recognition site peptide. CV-1 cells lack nuclear thyroid hormone Receptor, but express plasma membrane αVβ3; treatment of these cells with physiological concentrations of T4 activates the MAPK pathway, an effect inhibited by tetraiodothyroacetic acid, RGD peptide, and αVβ3 antibodies. Inhibitors of T4 binding to the integrin also block the MAPK-mediated proangiogenic action of T4. T4-induced phosphorylation of MAPK is inhibited by small interfering RNA knockdown of αV and β3. These findings suggest that T4 binds to αVβ...
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integrin alphavbeta3 contains a cell Surface Receptor site for thyroid hormone that is linked to activation of mitogen activated protein kinase and induction of angiogenesis
Endocrinology, 2005Co-Authors: Joel J Ergh, Lawrence Lansing, Seema Mohamed, Shaker A Mousa, Faith Davis, Paul J DavisAbstract:Integrin alpha(V)beta(3) is a heterodimeric plasma membrane protein whose several extracellular matrix protein ligands contain an RGD recognition sequence. This study identifies integrin alpha(V)beta(3) as a cell Surface Receptor for thyroid hormone [L-T(4) (T(4))] and as the initiation site for T(4)-induced activation of intracellular signaling cascades. Integrin alpha(V)beta(3) dissociably binds radiolabeled T(4) with high affinity, and this binding is displaced by tetraiodothyroacetic acid, alpha(V)beta(3) antibodies, and an integrin RGD recognition site peptide. CV-1 cells lack nuclear thyroid hormone Receptor, but express plasma membrane alpha(V)beta(3); treatment of these cells with physiological concentrations of T(4) activates the MAPK pathway, an effect inhibited by tetraiodothyroacetic acid, RGD peptide, and alpha(V)beta(3) antibodies. Inhibitors of T(4) binding to the integrin also block the MAPK-mediated proangiogenic action of T(4). T(4)-induced phosphorylation of MAPK is inhibited by small interfering RNA knockdown of alpha(V) and beta(3). These findings suggest that T(4) binds to alpha(V)beta(3) near the RGD recognition site and show that hormone-binding to alpha(V)beta(3) has physiological consequences.
