STRUCTURAL BIOLOGY OF DEATH-INDUCING RECEPTOR COMPLEXES

PhD student project 7
(Early Stage Researcher 7, ESR7)

STRUCTURAL BIOLOGY OF DEATH-INDUCING RECEPTOR COMPLEXES

Supervisor: Bert Janssen

Early Stage Researcher: Nadia Leloup

   

Aim:
In-depth characterisation of multi-domain proteins and multi-component complexes to determine the molecular mechanisms of signal triggering by pro-NGF through the p75-sortilin and p75-SorCS2 complexes.

Methodology:
ESR7 will use a combination of biophysical, structural and cellular analysis to detail the molecular mechanism underlying pro-NGF signal triggering leading to neuronal cell death or growth cone retraction. Extracellular protein segments that vary in size and, potentially, function will be expressed in HEK cells for biophysical and structural studies (collaboration UPE). Biophysical methods SPR, AUC, MALS and H/D Mass Spectrometry (collaboration Heck) will pinpoint and verify regions of importance for interaction and the stoichiometry of complexes. X-ray crystallography  and SAXS or EM (for samples that do not produce crystals) will reveal in detail the molecular basis of interaction and the changes that follow complex formation. Structure based mutants with altered functions will be tested in biophysical (e.g. SPR and AUC) and cellular assays addressing the full-length membrane spanning molecules to verify the interactions and oligomeric states of the receptors and co-receptors in the presence and absence of ligand (collaborations Killian, Lambris).

Collaborators:
Heck, Killian; Lambris (UPenn, USA); UPE

Key publications:

  1. B.J. Janssen, R.A. Robinson, F. Perez-Branguli, C.H. Bell, K.J. Mitchell, C. Siebold and E.Y. Jones. (2010) ‘Structural basis of semaphorin-plexin signalling.’ Nature 467 1118-1122
  2. B.J. Janssen, L. Gomes, R.I. Koning, D.I. Svergun, A.J. Koster, D.C. Fritzinger, C.W. Vogel and P. Gros. (2009) 'Insights into complement convertase formation based on the structure of the factor B-cobra venom factor complex.' Embo J 28 2469-2478
  3. B.J. Janssen, A. Christodoulidou, A. McCarthy, J.D. Lambris and P. Gros. (2006) 'Structure of C3b reveals conformational changes that underlie complement activity.' Nature 444 213-216