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PDBsum entry 4tpy
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References listed in PDB file
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Key reference
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Title
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High throughput screening using acoustic droplet ejection to combine protein crystals and chemical libraries on crystallization plates at high density.
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Authors
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E.Teplitsky,
K.Joshi,
D.L.Ericson,
A.Scalia,
J.D.Mullen,
R.M.Sweet,
A.S.Soares.
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Ref.
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J Struct Biol, 2015,
191,
49-58.
[DOI no: ]
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PubMed id
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Abstract
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We describe a high throughput method for screening up to 1728 distinct chemicals
with protein crystals on a single microplate. Acoustic droplet ejection (ADE)
was used to co-position 2.5nL of protein, precipitant, and chemicals on a
MiTeGen in situ-1 crystallization plate™ for screening by co-crystallization
or soaking. ADE-transferred droplets follow a precise trajectory which allows
all components to be transferred through small apertures in the microplate lid.
The apertures were large enough for 2.5nL droplets to pass through them, but
small enough so that they did not disrupt the internal environment created by
the mother liquor. Using this system, thermolysin and trypsin crystals were
efficiently screened for binding to a heavy-metal mini-library. Fluorescence and
X-ray diffraction were used to confirm that each chemical in the heavy-metal
library was correctly paired with the intended protein crystal. A fragment
mini-library was screened to observe two known lysozyme ligands using both
co-crystallization and soaking. A similar approach was used to identify
multiple, novel thaumatin binding sites for ascorbic acid. This technology
pushes towards a faster, automated, and more flexible strategy for high
throughput screening of chemical libraries (such as fragment libraries) using as
little as 2.5nL of each component.
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Secondary reference #1
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Title
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Hitting the target: fragment screening with acoustic in situ co-Crystallization of proteins plus fragment libraries on pin-Mounted data-Collection micromeshes.
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Authors
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X.Yin,
A.Scalia,
L.Leroy,
C.M.Cuttitta,
G.M.Polizzo,
D.L.Ericson,
C.G.Roessler,
O.Campos,
M.Y.Ma,
R.Agarwal,
R.Jackimowicz,
M.Allaire,
A.M.Orville,
R.M.Sweet,
A.S.Soares.
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Ref.
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Acta Crystallogr D Biol Crystallogr, 2014,
70,
1177-1189.
[DOI no: ]
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PubMed id
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