WNT-mediated signaling relay in stem cells and oncogenesis - from basic biology to applications

Targeting intrinsically disordered regions in the WNT pathway

Early stage researcher 10 (ESR10) project

Supervision: Assaf Friedler

Lab webpage:  http://chem.ch.huji.ac.il/~assaf

Aim:
Targeting intrinsically disordered regions in the WNT pathway for the development of anti-cancer drugs

Methodology:
ESR10 will target intrinsically disordered proteins of the WNT cascade including AXIN1 and APC by peptides derived from their protein interaction sites (private sector secondment: Pepscan). ESR10 will combine protein biochemistry, peptide arrays, peptide synthesis and biophysical methods. ESR10 will express and purify domains of these proteins and reveal which parts of the proteins are intrinsically disordered. Using peptide arrays, ESR10 will characterise the interactions of these proteins with their partners and identify the precise binding sites. This will be followed by quantifying the binding of the peptides to the partner protein and monitoring structural changes upon binding by CD and NMR (international secondment: Rüdiger). ESR10 will also test the effect of the peptides on the interactions and activity of the partner proteins in cells (collaboration Maurice).

Collaborators:
Rüdiger, Maurice, Pepscan, UPE

Project goals:
ESR10 will develop novel methodology to target IDRs and generate lead anti-cancer peptides that interfere with the WNT pathway. 

Risk assessment and contingency plans:
Problematic protein expression and purification will be assisted by UPE, which has a vast expertise; problems with cell penetration of the active peptides will be solved by adding a cell penetrating sequence.

Key publications:

1.  Gabizon R, Brandt T, Sukenik S, Lahav N, Lebendiker M, Shalev DE, Veprintsev D, Friedler A. (2012) Specific Recognition of p53 Tetramers by Peptides Derived from p53 Interacting Proteins.; PLoS One. 2012;7(5):e38060.

2.  Reingewertz TH, Shalev DE, Sukenik S, Blatt O, Rotem-Bamberger S, Lebendiker M, Larisch S, Friedler A. (2011) Mechanism of the Interaction between the Intrinsically Disordered C-Terminus of the Pro-Apoptotic ARTS Protein and the Bir3 Domain of XIAP. PLoS One. 6(9):e24655. Epub 2011 Sep 20

3.  Katz C. , Benyamini H. , Rotem S. , Lebendiker M. , Danieli T. , Dines, M. , Bronner, V. , Bravman, T. , Rüdiger S.  and Friedler A.  (2008): Molecular Basis of the Interaction Between the Anti-Apoptotic Bcl-2 Family Proteins and the Pro-Apoptotic Protein ASPP2; Proc. Natl. Acad. Sci. USA, 105(34):12277-82