DescriptionThe plasma-membrane of living cells is the major organelle for cellular signaling cascades. To warrant the diverse functions of a cell, communication between the cytoplasm and its organelles and the extracellular space is crucial. Thus, membrane-localized protein receptors activated by various messengers are key to transmit signals into the cell. Defective regulation of these signaling cascades may lead to cell death or uncontrolled proliferation. An important point is the interaction of these receptors with cytosolic proteins. To analyze such interactions we use micro-structured surfaces in combination with fluorescent microscopy (“micro-patterning assay”). This technique was developed to detect protein-protein interactions (Schwarzenbacher et al., 2008; Weghuber et al., 2010) and offers the possibility to measure and quantify also weak or short-lived interactions in-vivo. We visualized the interaction of the following membrane-receptors with their respective intracellular binding partners: EGF-receptor (Grb2), Insulin-receptor (IRS1-4), and ß1- and ß2 adrenergic receptors (Arrestin, G-proteins). Additionally, we analyzed the insulin-dependent transfer of Glucose-transporter 4 (Glut4) to the plasma-membrane. In a next step we started to determine the effects of various messengers (EGF, Insulin, Epinephrine,…) on the described interactions. By doing so we characterized variations in the interaction-properties of these interaction pairs upon application of messenger molecules. Since the micro-patterning assay is a robust technique to analyze a large number of cells within a short time, it is our endeavor to investigate the effects of further, medically relevant messenger molecules (secondary plant metabolites) on the interaction of the aforementioned signaling proteins.
|Period||26 Feb 2012|
|Event title||Biophysical Society Meeting: null|
|Location||San Diego, United States|