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PDBsum entry 2pu2
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References listed in PDB file
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Key reference
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Title
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Comprehensive mechanistic analysis of hits from high-Throughput and docking screens against beta-Lactamase.
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Authors
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K.Babaoglu,
A.Simeonov,
J.J.Irwin,
M.E.Nelson,
B.Feng,
C.J.Thomas,
L.Cancian,
M.P.Costi,
D.A.Maltby,
A.Jadhav,
J.Inglese,
C.P.Austin,
B.K.Shoichet.
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Ref.
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J Med Chem, 2008,
51,
2502-2511.
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PubMed id
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Abstract
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High-throughput screening (HTS) is widely used in drug discovery. Especially for
screens of unbiased libraries, false positives can dominate "hit lists"; their
origins are much debated. Here we determine the mechanism of every active hit
from a screen of 70,563 unbiased molecules against beta-lactamase using
quantitative HTS (qHTS). Of the 1,274 initial inhibitors, 95% were
detergent-sensitive and were classified as aggregators. Among the 70 remaining
were 25 potent, covalent-acting beta-lactams. Mass spectra, counter-screens, and
crystallography identified 12 as promiscuous covalent inhibitors. The remaining
33 were either aggregators or irreproducible. No specific reversible inhibitors
were found. We turned to molecular docking to prioritize molecules from the same
library for testing at higher concentrations. Of 16 tested, 2 were modest
inhibitors. Subsequent X-ray structures corresponded to the docking prediction.
Analog synthesis improved affinity to 8 microM. These results suggest that it
may be the physical behavior of organic molecules, not their reactivity, that
accounts for most screening artifacts. Structure-based methods may prioritize
weak-but-novel chemotypes in unbiased library screens.
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