9DOZ

Myocilin OLF oligomers

Summary for 9DOZ

• Entry DOI: 10.2210/pdb9doz/pdb
• Descriptor: Myocilin, CALCIUM ION, SODIUM ION, ... (4 entities in total)
• Functional Keywords: olf, cell adhesion
• Biological source: Homo sapiens (human)
• Total number of polymer chains: 1
• Total formula weight: 31260.05

Authors

Scelsi, H.F.,Huard, D.J.E.,Lieberman, R.L. (deposition date: 2024-09-20, release date: 2025-03-26, Last modification date: 2025-04-02)

Primary citation

Scelsi, H.F.,Close, E.G.S.,Huard, D.J.E.,Dunn, E.,Bogdanovic, N.,Mudiyanselage, S.H.W.,Grant, A.,Stagg, S.M.,Schmidt-Krey, I.,Van Horn, W.D.,Lieberman, R.L.
Detection of non-native species formed during fibrillization of the myocilin olfactomedin domain.
Protein Sci., 34:e70063-e70063, 2025

PubMed Abstract: Glaucoma is a group of neurodegenerative diseases that together are the leading cause of irreversible blindness worldwide. Myocilin-associated glaucoma is an inherited form of this disease, caused by intracellular aggregation of misfolded mutant myocilin. In vitro, the myocilin C-terminal olfactomedin domain (OLF), the relevant domain for glaucoma pathogenesis, can be driven to form amyloid-like fibrils under mild conditions. Here we characterize a species present during in vitro fibrillization. Purified OLF was subjected to fibrillization at concentrations required for downstream electron microscopy imaging and NMR spectroscopy. Additional biophysical techniques, including analytical ultracentrifugation and X-ray crystallography, were employed to further characterize the multicomponent mixture. Negative stain transmission electron microscopy (TEM) shows a non-native species reminiscent of known prefibrillar oligomers from other amyloid systems, NMR indicates a minor population of partially misfolded species is present in solution, and cryo-EM imaging shows two-dimensional protein arrays. The predominant soluble species remaining in solution after the fibril reaction is natively folded, as evidenced by X-ray crystallography. In summary, after incubating OLF under fibrillization-promoting conditions, there is a heterogeneous mixture consisting of soluble folded protein, mature amyloid-like fibrils, and partially misfolded intermediate species that at present belie additional molecular detail. The characterization of OLF fibrillar species illustrates the challenges associated with developing a comprehensive understanding of the fibrillization process for large, non-model amyloidogenic proteins.

PubMed: 40095382
DOI: 10.1002/pro.70063

Experimental method

X-RAY DIFFRACTION (1.454 Å)