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PDBsum entry 6nao
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Transferase/transferase inhibitor
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PDB id
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6nao
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References listed in PDB file
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Key reference
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Title
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Discovery of pf-06928215 as a high affinity inhibitor of cgas enabled by a novel fluorescence polarization assay.
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Authors
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J.Hall,
A.Brault,
F.Vincent,
S.Weng,
H.Wang,
D.Dumlao,
A.Aulabaugh,
D.Aivazian,
D.Castro,
M.Chen,
J.Culp,
K.Dower,
J.Gardner,
S.Hawrylik,
D.Golenbock,
D.Hepworth,
M.Horn,
L.Jones,
P.Jones,
E.Latz,
J.Li,
L.L.Lin,
W.Lin,
D.Lin,
F.Lovering,
N.Niljanskul,
R.Nistler,
B.Pierce,
O.Plotnikova,
D.Schmitt,
S.Shanker,
J.Smith,
W.Snyder,
T.Subashi,
J.Trujillo,
E.Tyminski,
G.Wang,
J.Wong,
B.Lefker,
L.Dakin,
K.Leach.
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Ref.
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PLoS One, 2017,
12,
e0184843.
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PubMed id
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Abstract
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Cyclic GMP-AMP synthase (cGAS) initiates the innate immune system in response to
cytosolic dsDNA. After binding and activation from dsDNA, cGAS uses ATP and GTP
to synthesize 2', 3' -cGAMP (cGAMP), a cyclic dinucleotide second messenger with
mixed 2'-5' and 3'-5' phosphodiester bonds. Inappropriate stimulation of cGAS
has been implicated in autoimmune disease such as systemic lupus erythematosus,
thus inhibition of cGAS may be of therapeutic benefit in some diseases; however,
the size and polarity of the cGAS active site makes it a challenging target for
the development of conventional substrate-competitive inhibitors. We report here
the development of a high affinity (KD = 200 nM) inhibitor from a low affinity
fragment hit with supporting biochemical and structural data showing these
molecules bind to the cGAS active site. We also report a new high throughput
cGAS fluorescence polarization (FP)-based assay to enable the rapid
identification and optimization of cGAS inhibitors. This FP assay uses
Cy5-labelled cGAMP in combination with a novel high affinity monoclonal antibody
that specifically recognizes cGAMP with no cross reactivity to cAMP, cGMP, ATP,
or GTP. Given its role in the innate immune response, cGAS is a promising
therapeutic target for autoinflammatory disease. Our results demonstrate its
druggability, provide a high affinity tool compound, and establish a high
throughput assay for the identification of next generation cGAS inhibitors.
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