G protein-coupled receptors : functional genomics, molecular pharmacology, structure-function studies.

Project Details

Description

The renin-angiotensin system is widely recognized as the most powerful hormonal system controlling sodium balance, body fluid volumes and arterial blood pressure. The effects of Ang-(1-7) and Ang IV are not all AT1/AT2 receptor-related and the existence of distinct receptors (respectively AT(1-7) and AT4) and even subtypes thereof has been proposed based on functional data (mostly on tissues) and radioligand binding (mostly on membranes).

Our project aims an unambiguous characterisation of the molecular and binding properties of such receptors and the nature of the activated signaling mechanisms within different tissues. Several strategies will be combined.
- The use of cell lines expressing receptors of interest. Radioligand binding and functional studies can be performed .
- Screening (HPLC and mass spectrometry) of proteolytic degradation of all peptides by each cell type and search for an optimal blockade of involved peptidases
- N-terminal tritiated radioligands will be used and synthesized.
- Measurement of rapid ‘second messenger’ responses (cAMP, inositol phosphates, [Ca2+]i (bioluminescence in aequorin-transfected cells), cGMP, protein-tyrosine phosphatase and Erk-2 activity, NO (microsensor-based), prostaglandin release, extracellular acidification (cytosensor)).
- photoaffinity labeling.

Using adequate tools and test conditions (see above), orphan GPCR’s will be screened for their interaction with these peptides.
AcronymDWTC121
StatusFinished
Effective start/end date1/01/0231/12/06

Flemish discipline codes

  • Basic sciences
  • Biological sciences

Keywords

  • cel-responsen
  • receptoren
  • proteolyse
  • cellijnen
  • radioligand binding
  • angiotensine fragmenten