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PDBsum entry 6asp
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Chaperone/inhibitor
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PDB id
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6asp
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References listed in PDB file
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Key reference
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Title
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Trifunctional high-Throughput screen identifies promising scaffold to inhibit grp94 and treat myocilin-Associated glaucoma.
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Authors
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D.J.E.Huard,
V.M.Crowley,
Y.Du,
R.A.Cordova,
Z.Sun,
M.O.Tomlin,
C.A.Dickey,
J.Koren,
L.Blair,
H.Fu,
B.S.J.Blagg,
R.L.Lieberman.
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Ref.
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ACS Chem Biol, 2018,
13,
933-941.
[DOI no: ]
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PubMed id
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Abstract
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Gain-of-function mutations within the olfactomedin (OLF) domain of myocilin
result in its toxic intracellular accumulation and hasten the onset of
open-angle glaucoma. The absence of myocilin does not cause disease; therefore,
strategies aimed at eliminating myocilin could lead to a successful glaucoma
treatment. The endoplasmic reticulum Hsp90 paralog Grp94 accelerates OLF
aggregation. Knockdown or pharmacological inhibition of Grp94 in cells
facilitates clearance of mutant myocilin via a non-proteasomal pathway. Here, we
expanded our support for targeting Grp94 over cytosolic paralogs Hsp90α and
Hsp90β. We then developed a high-throughput screening assay to identify new
chemical matter capable of disrupting the Grp94/OLF interaction. When applied to
a blind, focused library of 17 Hsp90 inhibitors, our miniaturized single-read in
vitro thioflavin T -based kinetics aggregation assay exclusively identified
compounds that target the chaperone N-terminal nucleotide binding site. In
follow up studies, one compound (2) decreased the extent of co-aggregation of
Grp94 with OLF in a dose-dependent manner in vitro, and enabled clearance of the
aggregation-prone full-length myocilin variant I477N in cells without inducing
the heat shock response or causing cytotoxicity. Comparison of the co-crystal
structure of compound 2 and another non-selective hit in complex with the
N-terminal domain of Grp94 reveals a docking mode tailored to Grp94 and explains
its selectivity. A new lead compound has been identified, supporting a targeted
chemical biology assay approach to develop a protein degradation-based therapy
for myocilin-associated glaucoma by selectively inhibiting Grp94.
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