Engineering a surrogate human heteromeric α/β glycine receptor orthosteric site exploiting the structural homology and stability of acetylcholine-binding protein

Alice Dawson, Paul Trumper, Juliana Oliveira de Souza, Holly Parker, Matthew J. Jones, Tim G. Hales, William N. Hunter (Lead / Corresponding author)

Research output: Contribution to journalArticle

2 Citations (Scopus)
93 Downloads (Pure)

Abstract

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.

Original languageEnglish
Pages (from-to)1014-1023
Number of pages10
JournalIUCrJ
Volume6
Issue numberPart 6
Early online date4 Sep 2019
DOIs
Publication statusPublished - 1 Nov 2019

Keywords

  • Acetylcholine-binding protein
  • crystal structures
  • glycine receptor
  • ligand-gated ion channel
  • nicotine
  • strychnine
  • tropisetron
  • acetylcholine-binding protein

Fingerprint Dive into the research topics of 'Engineering a surrogate human heteromeric α/β glycine receptor orthosteric site exploiting the structural homology and stability of acetylcholine-binding protein'. Together they form a unique fingerprint.

  • Cite this