1. The Folding of WW Domain FBP28 (Formin binding protein): C-term N-term Xavier Periole

2. WW domains • Two highly conserved tryptophans • ~ 35-45 aa. • Non-catalytic signaling proteins (protein-protein recognition) • >200 non-redundant proteins (2003) • Bind proline-rich sequences. • 6 classes (sequence preference).

3. WW Domain FBP28 • WW domains: • Fast and reversible folding • Highly conserved structure • Rapid formation of amyloid fibres (Protein Conformational Disorder) • Collaboration with Guy Lippens (NMR-Lille) • WW Domain FBP28 (Formin binding protein): • Fastest folding of the family (10s of s) • The fold is conserved upon mutations • Temp, mutations and sequence truncation modulate a three- to two-state folding pathway • The N- (res 1-5) and C-termini (res 34-37) are not required: 28 residues Xavier Periole

4. WW Domain FBP28 Very stable and highly conserved slightly curved three-stranded antiparallel ß-sheet C-term N-term Xavier Periole

5. WW Domain FBP28 • 38 Replicas • T: from 282 to 498K • in water • GROMOS96 FF 38*200ns=7.6ms Xavier Periole

6. WW Domain FBP28 RMSD from the X-ray structure T=280K T=500K Xavier Periole

7. WW Domain FBP28 Structures with minimum deviation from the X-ray X-ray rmsd 0.22 nm rmsd 0.14 nm Xavier Periole

8. WW Domain FBP28 RMSD from the X-ray structure Xavier Periole

9. WW Domain FBP28 rmsd vs. Rgyr Xavier Periole

10. WW Domain FBP28 Comparison with the Native State rmsd vs. Sec. Struct. Xavier Periole

11. WW Domain FBP28 rmsd vs. native contacts Xavier Periole

12. The Real Case Build the Transition State • Characterize the limits native/unfolded states • Build the intermediary state • Unfolding Simulation • Use F-values as restraints in REMD Xavier Periole

13. The Real Case REMD from the native state: Unfolding simulations Xavier Periole

14. The Real Case F-values as restraints in MD: b) a) rmsd: 4.4 1.3Å rmsd (Å) A=0 A=1 Mutation does not affect the TSE Mutation does affect the TSE Xavier Periole