Abstract
The recent structural elucidation of ex vivo Drosophila Orb2 fibrils revealed a novel amyloid formed by interdigitated Gln and His residue side chains belonging to the prion-like domain. However, atomic-level details on the conformational transitions associated with memory consolidation remain unknown. Here, we have characterized the nascent conformation and dynamics of the prion-like domain (PLD) of Orb2A using a nonconventional liquid-state NMR spectroscopy strategy based on 13C detection to afford an essentially complete set of 13Cα, 13Cβ, 1Hα and backbone 13CO and 15N assignments. At pH 4, where His residues are protonated, the PLD is disordered and flexible, except for a partially populated α-helix spanning residues 55-60. At pH 7, in contrast, His residues are predominantly neutral and the Q/H segments adopt minor populations of helical structure, show decreased mobility and start to self-associate. At pH 7, the His residues also bind Zn++, which promotes further association. These findings represent a remarkable case of structural plasticity, based on which an updated model for Orb2A functional amyloidogenesis is advanced.
Highlights
· The Orb2 prion like domain that forms the structures related to memory consolidation is studied by solution NMR.
· The amyloidogenic Q/H-rich stretch is disordered and flexible at low pH.
· Residues 55-60 form a partly populated α-helix at pH 4.
· At pH 7, the Q/H-rich segment also adopts a low population of α-helix and rigidifies.
· Zn++ binding induces associative changes in the Orb2 prion-like domain.
Competing Interest Statement
The authors have declared no competing interest.
Footnotes
The revised manuscript includes some additional grant information. Everything else is the same.
Abbreviations Used
- CPEB
- Cytoplasmic Polyadenylation Element Binding protein.
- Orb2A
- The Drosophila protein encoded by the oo18 RNA-binding (orb) gene, predominantly neuronal isoform 2A.
- IDR
- Intrinsically Disordered Region (Residues 1-292 of Orb2A)
- NMR
- Nuclear Magnetic Resonance
- PLD
- Prion-Like Domain (Residues 1-68 of Orb2A)