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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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PDB id:
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| Name: |
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Signaling protein
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Title:
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Crystal structure of mutant achbp in complex with strychnine (t53f, q74r, y110a, i135s, g162e)
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Structure:
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Soluble acetylcholine receptor. Chain: a, b, c, d, e, f, g, h, i, j, k, l, m, n, o. Engineered: yes. Mutation: yes
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Source:
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Aplysia californica. California sea hare. Organism_taxid: 6500. Expressed in: spodoptera frugiperda. Expression_system_taxid: 7108. Expression_system_variant: sf9
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Resolution:
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3.20Å
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R-factor:
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0.190
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R-free:
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0.231
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Authors:
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A.Dawson,W.N.Hunter,J.O.De Souza,P.Trumper
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Key ref:
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A.Dawson
et al.
(2019).
Engineering a surrogate human heteromeric α/β glycine receptor orthosteric site exploiting the structural homology and stability of acetylcholine-binding protein.
IUCrJ,
6,
1014-1023.
PubMed id:
DOI:
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Date:
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22-Jun-17
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Release date:
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01-Aug-18
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PROCHECK
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Headers
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References
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Q8WSF8
(Q8WSF8_APLCA) -
Soluble acetylcholine receptor from Aplysia californica
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Seq:
Struc:
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236 a.a.
206 a.a.*
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Key: |
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PfamA domain |
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Secondary structure |
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*
PDB and UniProt seqs differ
at 7 residue positions (black
crosses)
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DOI no:
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IUCrJ
6:1014-1023
(2019)
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PubMed id:
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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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A.Dawson,
P.Trumper,
J.O.de Souza,
H.Parker,
M.J.Jones,
T.G.Hales,
W.N.Hunter.
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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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Links
