title: Application of fragment-based drug discovery to membrane proteins: identification of ligands of the integral membrane enzyme DsbB.
abstract: Membrane proteins are important pharmaceutical targets, but they pose significant challenges for fragment-based drug discovery approaches. Here, we present the first successful use of biophysical methods to screen for fragment ligands to an integral membrane protein. The Escherichia coli inner membrane protein DsbB was solubilized in detergent micelles and lipid bilayer nanodiscs. The solubilized protein was immobilized with retention of functionality and used to screen 1071 drug fragments for binding using target immobilized NMR Screening. Biochemical and biophysical validation of the eight most potent hits revealed an IC(50) range of 7-200 microM. The ability to insert a broad array of membrane proteins into nanodiscs, combined with the efficiency of TINS, demonstrates the feasibility of finding fragments targeting membrane proteins.
authors.affiliation: Leiden Institute of Chemistry, Leiden University, Leiden 2300RA, The Netherlands.
authors.forename: Virginie
authors.initials: V
authors.lastname: Früh
authors.forename: Yunpeng
authors.initials: Y
authors.lastname: Zhou
authors.forename: Dan
authors.initials: D
authors.lastname: Chen
authors.forename: Caroline
authors.initials: C
authors.lastname: Loch
authors.forename: Eiso
authors.initials: E
authors.lastname: Ab
authors.forename: Yelena N
authors.initials: YN
authors.lastname: Grinkova
authors.forename: Herman
authors.initials: H
authors.lastname: Verheij
authors.forename: Stephen G
authors.initials: SG
authors.lastname: Sligar
authors.forename: John H
authors.initials: JH
authors.lastname: Bushweller
authors.forename: Gregg
authors.initials: G
authors.lastname: Siegal
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D055808:Drug Discovery / Q000379:methods*; D004926:Escherichia coli; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D008823:Micelles; D036103:Nanotechnology; D011485:Protein Binding; D055550:Protein Stability; D015203:Reproducibility of Results; D012995:Solubility
chemical_list: D001426:Bacterial Proteins; C079503:DsbB protein, Bacteria; D055767:Immobilized Proteins; D008024:Ligands; D008565:Membrane Proteins; D008823:Micelles
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