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TOPIC: Biospectroscopy.

Alterations of δ-opioid receptor mobility and function by acute and chronic depletion of plasma membrane cholesterol content of live cells

2017-2019

17-05903S, Grant Agency of the Czech Republic (associated contractor - project leader is affiliated with another institution)

Detailed analysis of the effect of acute and chronic depletion of cell cholesterol level on δ-opioid receptor mobility and function in plasma membrane of model cell lines. For this aim, HEK293 and NG108-15 cells transiently expressing δ-OR-eCFP and δ-OR-eYFP will be analyzed by  advanced techniques of fluorescence microscopy FRAP, RICS, FCS, TIRF and 2PPM. Subsequently, alterations of δ-OR mobility will be correlated with alterations of agonist ligand binding, and function of δ-OR measured by agonist-stimulated [35S]GTPγS binding. These experiments will be performed in HEK293 cells stably expressing FLAG-δ-OR and NG108-15 cells. "Antibody feeding" method will be used to distinguish internalized and cell surface pools of δ-OR. Functional studies of δ-OR will be performed in isolated plasma membranes and detergent-untreated, low-density membrane domains. Project will extend the insight into the toxic side-effects of statin-based therapies and bring new knowledge about correlation between lateral mobility of δ-OR in plasma membrane of live cells and its functional significance.

    Mgr. SĂťKORA Jan, Ph. D.

    Department: Department of Biophysical Chemistry
    Phone: (+420) 26605 3142, 3484, 3187
    Fax: (+420) 286582307
    E-mail: jan.sykoraatjh-inst.cas.cz
    WWW: http://www.jh-inst.cas.cz/~fluorescence/
    Room: 212, 214, 218


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