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PDBsum entry 5oal
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Signaling protein
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PDB id
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5oal
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References listed in PDB file
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Key reference
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Title
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Engineering a surrogate human heteromeric α/β glycine receptor orthosteric site exploiting the structural homology and stability of acetylcholine-Binding protein.
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Authors
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A.Dawson,
P.Trumper,
J.O.De souza,
H.Parker,
M.J.Jones,
T.G.Hales,
W.N.Hunter.
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Ref.
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IUCrJ, 2019,
6,
1014-1023.
[DOI no: ]
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PubMed id
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Abstract
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Protein-engineering methods have been exploited to produce a surrogate system
for the extracellular neurotransmitter-binding site of a heteromeric human
ligand-gated ion channel, the glycine receptor. This approach circumvents two
major issues: the inherent experimental difficulties in working with a
membrane-bound ion channel and the complication that a heteromeric assembly is
necessary to create a key, physiologically relevant binding site. Residues that
form the orthosteric site in a highly stable ortholog, acetylcholine-binding
protein, were selected for substitution. Recombinant proteins were prepared and
characterized in stepwise fashion exploiting a range of biophysical techniques,
including X-ray crystallography, married to the use of selected chemical probes.
The decision making and development of the surrogate, which is termed a
glycine-binding protein, are described, and comparisons are provided with
wild-type and homomeric systems that establish features of molecular recognition
in the binding site and the confidence that the system is suited for use in
early-stage drug discovery targeting a heteromeric α/β glycine receptor.
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