Skip to main content

Dissecting pathogen recognition complexes of Toll-like receptors: hunting for new co-receptors and ligands

Objective

Toll-like receptors (TLRs) are trans-membrane proteins that recognize pathogen associated molecular patterns (PAMPs) and subsequently induce an intracellular signaling cascade that triggers the activation of transcription factors which initiate the expression of host defense proteins. Ten human, structurally highly conserved TLRs have been identified and assigned to structurally very dissimilar ligands. The complexity of a PAMP-TLR interface is best understood for the recognition of Lipopolysaccharide (LPS) by a LBP-Cd14,TLR4-Md2 receptor complex. The central hypothesis of this project is that many further ligand specific co-receptors and co-ligands remain to be discovered. I propose to systematically search for endosomal TLR receptor complexes required for the innate immune response to viral infection. Therefore, an interdisciplinary strategy combining molecular biology, cell biology, chemistry and proteomics will be employed. Recombinant ectodomains will be purified and used for monoclonal antibody production and affinity chromatography with macrophage lysates to fish cellular binding proteins. Following pre-enrichment of endosomes, endogenous TLRs will be immunoprecipitated from macrophages. Moreover, ligand affinity chromatography based proteomics using synthetic analogues like Poly(I:C) or the small compound imiquimod and tandem affinity purification of TLRs provide additional methods for complex purifications. Following identification of complexes by mass spectrometry, interactors will be validated and further characterized using cell biological, biochemical and immunological methods. This project will help to understand the process of viral pathogen recognition by the innate immune system and can provide novel drug targets for the tuning of innate immune response.

Field of science

  • /natural sciences/biological sciences/cell biology
  • /natural sciences/biological sciences/molecular biology
  • /natural sciences/biological sciences/biochemistry/biomolecules/proteins
  • /medical and health sciences/basic medicine/immunology
  • /natural sciences/biological sciences/biochemistry/biomolecules/proteins/proteomics
  • /natural sciences/chemical sciences/analytical chemistry/mass spectrometry

Call for proposal

FP7-PEOPLE-2007-2-1-IEF
See other projects for this call

Funding Scheme

MC-IEF - Intra-European Fellowships (IEF)

Coordinator

CEMM - FORSCHUNGSZENTRUM FUER MOLEKULARE MEDIZIN GMBH
Address
Lazarettgasse 14 Akh Bt 25.3
1090 Wien
Austria
Activity type
Private for-profit entities (excluding Higher or Secondary Education Establishments)
EU contribution
€ 167 960,25
Administrative Contact
Giulio Superti-Furga (Prof.)