title: Design, synthesis and in vitro evaluation of novel bivalent S-adenosylmethionine analogues.
abstract: In optimal cases, bivalent ligands can bind with exceptionally high affinity to their protein targets. However, designing optimised linkers, that orient the two binding groups perfectly, is challenging, and yet crucial in both fragment-based ligand design and in the discovery of bisubstrate enzyme inhibitors. To further our understanding of linker design, a series of novel bivalent S-adenosylmethionine (SAM) analogues were designed with the aim of interacting with the MetJ dimer in a bivalent sense (1:1 ligand/MetJ dimer). A range of ligands was synthesised and analyzed for ability to promote binding of the Escherichia coli methionine repressor, MetJ, to its operator DNA. Binding of bivalent SAM analogues to the MetJ homodimer in the presence of operator DNA was evaluated by fluorescence anisotropy and the effect of linker length and structure was investigated. The most effective bivalent ligand identified had a flexible linker, and promoted the DNA-protein interaction at 21-times lower concentration than the corresponding monovalent control compound.
authors.affiliation: School of Chemistry, University of Leeds, Leeds LS2 9JT, UK.
authors.forename: Catherine
authors.initials: C
authors.lastname: Joce
authors.forename: Rebecca
authors.initials: R
authors.lastname: White
authors.forename: Peter G
authors.initials: PG
authors.lastname: Stockley
authors.forename: Stuart
authors.initials: S
authors.lastname: Warriner
authors.forename: W Bruce
authors.initials: WB
authors.lastname: Turnbull
authors.forename: Adam
authors.initials: A
authors.lastname: Nelson
mesh_terms: D016880:Anisotropy; D001426:Bacterial Proteins / Q000737:chemistry; D002626:Chemistry, Pharmaceutical / Q000379:methods; D004247:DNA / Q000737:chemistry; D019281:Dimerization; D015195:Drug Design; D004926:Escherichia coli / Q000378:metabolism; D008024:Ligands; D008956:Models, Chemical; D008968:Molecular Conformation; D011485:Protein Binding; D012097:Repressor Proteins / Q000737:chemistry; D012436:S-Adenosylmethionine / Q000031:analogs & derivatives / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000379:methods
chemical_list: D001426:Bacterial Proteins; D008024:Ligands; D012097:Repressor Proteins; C060486:methionine repressor protein, Bacteria; D012436:S-Adenosylmethionine; D004247:DNA