CHAPERONE BINDING MOTIFS IN DISORDERED PROTEINS

PhD student project 10
(Early Stage Researcher 10, ESR 10)

CHAPERONE BINDING MOTIFS IN DISORDERED PROTEINS

Supervision: Stefan Rüdiger

Early Stage Researcher: Tania Morán Luengo
 

   

Aim:
The aim of this project is to identify the binding site of the chaperone Hsp90 in one of its natural substrate proteins.

Methodology:
ESR10 will analyse peptide arrays to identify the binding motif in its natural substrates. ESR1 will further design peptide and protein fragments, also based on a structural model of an Hsp90-substrate complex that the Rüdiger group very recently obtained by NMR and SAXS. ESR10 will learn peptide synthesis (International secondment: Friedler). ESR10 will also analyse the interaction of posttranslationally modified peptides and fragments to test how those alterations affects Hsp90-substrate interaction. ESR10 will also produce longer disordered proteins (UPE), fragments and peptides to identify a general motif in substrates. The Hsp90-substrate interaction will be quantified by fluorescence and NMR spectroscopy, profiting from a recent technical breakthrough of the Rüdiger groups to analyse Hsp90 by NMR (collaboration Boelens). The stoichiometry of the complexes will be determined by native Mass Spectrometry (collaboration Heck). ESR10 will also explore whether Hsp90-peptide complexes can be crystallised (collaboration Janssen).

Collaborators:
Boelens, Janssen, Heck, Friedler, UPE

Project goals:
ESR10 will identify for the first time a binding site of the Hsp90 chaperone in substrates, will abstract a binding motif and will get mechanistic insights into how Hsp90 interferes with aggregation-linked diseases (e.g. Alzheimer’s Disease).

Key publications:

  1. G.E. Karagöz, A.M.S. Duarte, J. Ippel, C. Uetrecht, T. Sinnige, M. van Rosmalen , J. Hausmann, A.J.R. Heck, R. Boelens and S.G.D Rüdiger ‘The N-terminal domain of human Hsp90 triggers binding to the co-chaperone p23’, Proc Natl Acad Sci USA, 108, 580 (2011).
  2. D.P. Minde, Z. Anvarian, S.G.D. Rüdiger# and Maurice, MM# (2011) # joint corresponding authors. ‘Messing up disorder: How do missense mutations in the tumor suppressor protein APC lead to cancer?’ Mol Cancer 10, 101 (2011). Marked “highly accessed” by publisher.
  3. T. Didenko, R. Boelens and S.G.D. Rüdiger (2011). ‘3D DOSY–TROSY to determine the translational diffusion coefficient of large protein complexes.’ PEDS 24, 99-103.

 

Cutting edge NMR spectroscopy to study the 170 kDa Hsp90 dimer.