Title | Ligands for glaucoma-associated myocilin discovered by a generic binding assay. |
Publication Type | Journal Article |
Year of Publication | 2014 |
Authors | Orwig, SD, Chi, PV, Du, Y, Hill, SE, Cavitt, MA, Suntharalingam, A, Turnage, KC, Dickey, CA, France, S, Fu, H, Lieberman, RL |
Journal | ACS Chem Biol |
Volume | 9 |
Issue | 2 |
Pagination | 517-25 |
Date Published | 2014 Feb 21 |
ISSN | 1554-8937 |
Keywords | Cytoskeletal Proteins, Drug Evaluation, Preclinical, Extracellular Matrix Proteins, Eye Proteins, Glaucoma, Open-Angle, Glycoproteins, Humans, Ligands, Models, Molecular, Mutation, Protein Binding, Protein Structure, Tertiary, Small Molecule Libraries, Structure-Activity Relationship, Surface Plasmon Resonance |
Abstract | Mutations in the olfactomedin domain of myocilin (myoc-OLF) are the strongest link to inherited primary open angle glaucoma. In this recently identified protein misfolding disorder, aggregation-prone disease variants of myocilin hasten glaucoma-associated elevation of intraocular pressure, leading to vision loss. Despite its well-documented pathogenic role, myocilin remains a domain of unknown structure or function. Here we report the first small-molecule ligands that bind to the native state of myoc-OLF. To discover these molecules, we designed a general label-free, mix-and-measure, high throughput chemical assay for restabilization (CARS), which is likely readily adaptable to discover ligands for other proteins. Of the 14 hit molecules identified from screening myoc-OLF against the Sigma-Aldrich Library of Pharmacologically Active Compounds using CARS, surface plasmon resonance binding studies reveal three are stoichiometric ligand scaffolds with low micromolar affinity. Two compounds, GW5074 and apigenin, inhibit myoc-OLF amyloid formation in vitro. Structure-activity relationship-based soluble derivatives reduce aggregation in vitro as well as enhance secretion of full-length mutant myocilin in a cell culture model. Our compounds set the stage for a new chemical probe approach to clarify the biological function of wild-type myocilin and represent lead therapeutic compounds for diminishing intracellular sequestration of toxic mutant myocilin. |
DOI | 10.1021/cb4007776 |
Alternate Journal | ACS Chem. Biol. |
PubMed ID | 24279319 |
PubMed Central ID | PMC3944078 |
Grant List | R01 EY021205 / EY / NEI NIH HHS / United States R01 EY024232 / EY / NEI NIH HHS / United States R01 NS073899 / NS / NINDS NIH HHS / United States R01EY021205 / EY / NEI NIH HHS / United States R01NS073899 / NS / NINDS NIH HHS / United States |
Ligands for glaucoma-associated myocilin discovered by a generic binding assay.
Submitted by enguyen7 on December 30, 2015 - 10:37am