The kinetic analysis of ligand receptor interactions suggests that receptors and G proteins exist in rapidly coupling and slowly coupling pools in membrane surfaces. To determine whether receptors in the slowly coupling pools were physically precoupled to G proteins or coupled rapidly, we have examined a number of peptides for their ability to interfere with receptor-G protein coupling in broken cell preparations. These peptides represent portions of receptors and G proteins in regions alledged to mediate coupling during signal transduction. Although these peptides have been shown to affect the interactions between the purified proteins, they do not seem to affect the interactions of the receptors and G protein in the cell membranes. This observation leads us to suggest that the receptors and G proteins are physically precoupled, supporting the model derived from kinetic analysis. The next phase of the project will involve a description of the underlying biochemistry of signal transduction. The kinetic methods will combine rapid mix flow cytometry with biochemisty to evaluate the evolution of receptors among different macromolecular complexes. These receptor complexes will be isolated by immunopurification of epitope tagged receptor and the transduction molecules in the comples will be identified. We will attempt to develop a mathematial model from the kinetic methods to account for the biochemical steps in receptor activation and desensitization.
| Frumkin, Jesse P; Patra, Biranchi N; Sevold, Anthony et al. (2016) The interplay between chromosome stability and cell cycle control explored through gene-gene interaction and computational simulation. Nucleic Acids Res 44:8073-85 |
| Ai, Ye; Sanders, Claire K; Marrone, Babetta L (2013) Separation of Escherichia coli bacteria from peripheral blood mononuclear cells using standing surface acoustic waves. Anal Chem 85:9126-34 |
| Sanders, Claire K; Mourant, Judith R (2013) Advantages of full spectrum flow cytometry. J Biomed Opt 18:037004 |
| Cushing, Kevin W; Piyasena, Menake E; Carroll, Nick J et al. (2013) Elastomeric negative acoustic contrast particles for affinity capture assays. Anal Chem 85:2208-15 |
| Johnson, Leah M; Gao, Lu; Shields IV, C Wyatt et al. (2013) Elastomeric microparticles for acoustic mediated bioseparations. J Nanobiotechnology 11:22 |
| Micheva-Viteva, Sofiya N; Shou, Yulin; Nowak-Lovato, Kristy L et al. (2013) c-KIT signaling is targeted by pathogenic Yersinia to suppress the host immune response. BMC Microbiol 13:249 |
| Piyasena, Menake E; Austin Suthanthiraraj, Pearlson P; Applegate Jr, Robert W et al. (2012) Multinode acoustic focusing for parallel flow cytometry. Anal Chem 84:1831-9 |
| Austin Suthanthiraraj, Pearlson P; Piyasena, Menake E; Woods, Travis A et al. (2012) One-dimensional acoustic standing waves in rectangular channels for flow cytometry. Methods 57:259-71 |
| Vuyisich, Momchilo; Sanders, Claire K; Graves, Steven W (2012) Binding and cell intoxication studies of anthrax lethal toxin. Mol Biol Rep 39:5897-903 |
| Chaudhary, Anu; Ganguly, Kumkum; Cabantous, Stephanie et al. (2012) The Brucella TIR-like protein TcpB interacts with the death domain of MyD88. Biochem Biophys Res Commun 417:299-304 |
Showing the most recent 10 out of 240 publications
