Evidence for S 331 -G-S-L within the amyloid core of myocilin olfactomedin domain fibrils based on low-resolution 3D solid-state NMR spectra

• Published in: bioRxiv
• Main Authors: Saccuzzo, Emily G, Robang, Alicia S, Gao, Yuan, Chen, Bo, Lieberman, Raquel L, Paravastu, Anant K
• Format: Journal Article
• Language: English
• Published: United States Cold Spring Harbor Laboratory 09.08.2024
• Subjects: solid-state nuclear magnetic resonance spectroscopy; Protein misfolding; proteostasis; amyloid fibrils; glaucoma
• ISSN: 2692-8205
• DOI: 10.1101/2024.08.09.606901

Myocilin-associated glaucoma is a protein-conformational disorder associated with formation of a toxic amyloid-like aggregate. Numerous destabilizing single point variants, distributed across the myocilin olfactomedin β-propeller (OLF, myocilin residues 245-504, 30 kDa) are associated with accelerated disease progression. , wild type (WT) OLF can be promoted to form thioflavin T (ThT)-positive fibrils under mildly destabilizing (37°C, pH 7.2) conditions. Consistent with the notion that only a small number of residues within a protein are responsible for amyloid formation, 3D C- C solid-state NMR spectra show that OLF fibrils are likely to be composed of only about one third of the overall sequence. Here, we probe the residue composition of fibrils formed from purified full-length OLF. We were able to make sequential assignments consistent with the sequence S -G-S-L . This sequence appears once within a previously identified amyloid-prone region (P1, G AVVYSGSLYFQ) internal to OLF. Since nearly half of the pairs of adjacent residues (di-peptides) in OLF occur only once in the primary structure and almost all the 3-residue sequences (tri-peptides) are unique, remarkably few sequential assignments are necessary to uniquely identify specific regions of the amyloid core. This assignment approach could be applied to other systems to expand our molecular comprehension of how folded proteins undergo fibrillization.