Impact of Lysine Acetylation Mutations on the Structure of Full-Length Tau Fibrils.

Abstract

The tau protein aggregates into amyloid fibrils in Alzheimer's disease and other neurodegenerative diseases. In these tauopathies, tau is decorated with posttranslational modifications, including phosphorylation and acetylation, suggesting that these modifications may cause tau to aggregate into specific pathological structures. Here, we investigate how pseudoacetylation of three lysine residues, K311Q, K321Q, and K369Q, affects the fibrilization and fibril structure of full-length four-repeat tau. These acetyl mimics are in addition to four phospho-mimetic glutamate mutations at the PHF1 epitope (4E tau). The joint mutant 4E3Q tau formed well-ordered amyloid fibrils without anionic cofactors. The 4E3Q tau fibrils lack twists, preventing structure determination by cryoelectron microscopy and necessitating characterization by solid-state NMR. ¹³C and ¹⁵N chemical shifts indicate that pseudoacetylation caused the protein to adopt a distinct fold from the parent 4E tau fibrils: the rigid core contains β-strands between R2 and R4 repeats and near the end of the C-terminal domain. Importantly, the C-terminal half of the R3 repeat containing the K321Q mutation is disordered, in qualitative contrast with 4E tau. Chemical shifts indicate that these structural changes likely result from the disruption of salt bridges between lysine and aspartate residues. 4E3Q tau contains an immobilized R2, which differs from that of AD tau. These results provide insights into the impact of acetylation on tau fibrilization and fibril structure and suggest that acetylation of these three lysine residues in AD may occur after the formation of the paired-helical filament structure.