title: Identification of Ligand-Receptor Interactions: Ligand Molecular Arrays, SPR and NMR Methodologies.
abstract: Despite many years of research into bacterial chemotaxis, the only well characterized system to date is that of E. coli. Even for E. coli, the direct ligand binding had been fully characterized only for aspartate and serene receptors Tar and Tsr. In 30 years since, no other direct receptor-ligand interaction had been described for bacteria, until the characterization of the C. jejuni aspartate and multiligand receptors (Hartley-Tassell et al. Mol Microbiol 75:710-730, 2010). While signal transduction components of many sensory pathways have now been characterized, ligand-receptor interactions remain elusive due to paucity of high-throughput screening methods. Here, we describe the use of microarray screening we developed to identify ligands, surface plasmon resonance, and saturation transfer difference nuclear magnetic resonance (STD-NMR) we used to verify the hits and to determine the affinity constants of the interactions, allowing for more targeted verification of ligands with traditional chemotaxis and in vivo assays described in Chapter 13 .
authors.affiliation: Institute for Glycomics, Griffith University, Gold Coast Campus, Gold Coast, Australia.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Day
authors.affiliation: Institute for Glycomics, Griffith University, Gold Coast Campus, Gold Coast, Australia.
authors.forename: Lauren E
authors.initials: LE
authors.lastname: Hartley-Tassell
authors.affiliation: Institute for Glycomics, Griffith University, Gold Coast Campus, Gold Coast, Australia. v.korolik@griffith.edu.au.
authors.forename: Victoria
authors.initials: V
authors.lastname: Korolik
mesh_terms: D000596:Amino Acids / Q000378:metabolism / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D016123:Campylobacter jejuni / Q000235:genetics* / Q000378:metabolism; D002633:Chemotaxis / Q000235:genetics; D003837:Deoxy Sugars / Q000378:metabolism / Q000494:pharmacology; D004926:Escherichia coli / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D057166:High-Throughput Screening Assays; D056656:Lab-On-A-Chip Devices; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D046208:Microchip Analytical Procedures; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism*; D012266:Ribose / Q000378:metabolism / Q000494:pharmacology; D054852:Small Molecule Libraries / Q000378:metabolism / Q000494:pharmacology*; D020349:Surface Plasmon Resonance
chemical_list: D000596:Amino Acids; D001426:Bacterial Proteins; D003837:Deoxy Sugars; D008024:Ligands; D011956:Receptors, Cell Surface; D054852:Small Molecule Libraries; D012266:Ribose
keywords: Chemotaxis receptors; High-throughput screening; Microarray ligand identification; NMR; SPR