ausblenden:
Schlagwörter:
TRANSMISSION ELECTRON-MICROSCOPY; SOLID-STATE NMR; BETA-SOLENOID
PROTEINS; URE2P PRION FILAMENTS; HUNTINGTONS-DISEASE; POLYGLUTAMINE;
PARALLEL; IMAGE; FORM; ORGANIZATION
Zusammenfassung:
The established correlation between neurodegenerative disorders and intracerebral deposition of polyglutamine aggregates motivates attempts to better understand their fibrillar structure. We designed polyglutamines with a few lysines inserted to overcome the hindrance of extreme insolubility and two D-lysines to limit the lengths of beta-strands. One is 33 amino acids long (PolyQKd-33) and the other has one fewer glutamine (PolyQKd-32). Both form well-dispersed fibrils suitable for analysis by electron microscopy. Electron diffraction confirmed cross-beta structures in both fibrils. Remarkably, the deletion of just one glutamine residue from the middle of the peptide leads to substantially different amyloid structures. PolyQKd-32 fibrils are consistently 10-20% wider than PolyQKd-33, as measured by negative staining, cryo-electron microscopy, and scanning transmission electron microscopy. Scanning transmission electron microscopy analysis revealed that the PolyQKd-32 fibrils have 50% higher mass-per-length than PolyQKd-33. This distinction can be explained by a superpleated beta-structure model for PolyQKd-33 ;and a model with two beta-solenoid protofibrils for PolyQKd-32. These data provide evidence for beta-arch-containing structures in polyglutamine fibrils and open future possibilities for structure-based drug design.
