title: Label free fragment screening using surface plasmon resonance as a tool for fragment finding - analyzing parkin, a difficult CNS target.
abstract: Surface Plasmon Resonance (SPR) is rarely used as a primary High-throughput Screening (HTS) tool in fragment-based approaches. With SPR instruments becoming increasingly high-throughput it is now possible to use SPR as a primary tool for fragment finding. SPR becomes, therefore, a valuable tool in the screening of difficult targets such as the ubiquitin E3 ligase Parkin. As a prerequisite for the screen, a large number of SPR tests were performed to characterize and validate the active form of Parkin. A set of compounds was designed and used to define optimal SPR assay conditions for this fragment screen. Using these conditions, more than 5000 pre-selected fragments from our in-house library were screened for binding to Parkin. Additionally, all fragments were simultaneously screened for binding to two off target proteins to exclude promiscuous binding compounds. A low hit rate was observed that is in line with hit rates usually obtained by other HTS screening assays. All hits were further tested in dose responses on the target protein by SPR for confirmation before channeling the hits into Nuclear Magnetic Resonance (NMR) and other hit-confirmation assays.
authors.affiliation: Elan Pharmaceuticals, South San Francisco, California, USA. Karin.Regnstrom@gmail.com
authors.forename: Karin
authors.initials: K
authors.lastname: Regnström
authors.forename: Jiangli
authors.initials: J
authors.lastname: Yan
authors.forename: Lan
authors.initials: L
authors.lastname: Nguyen
authors.forename: Kari
authors.initials: K
authors.lastname: Callaway
authors.forename: Yanli
authors.initials: Y
authors.lastname: Yang
authors.forename: Linnea
authors.initials: L
authors.lastname: Diep
authors.forename: Weimei
authors.initials: W
authors.lastname: Xing
authors.forename: Anirban
authors.initials: A
authors.lastname: Adhikari
authors.forename: Paul
authors.initials: P
authors.lastname: Beroza
authors.forename: Roy K
authors.initials: RK
authors.lastname: Hom
authors.forename: Brigit
authors.initials: B
authors.lastname: Riley
authors.forename: Don
authors.initials: D
authors.lastname: Rudolph
authors.forename: Michael F
authors.initials: MF
authors.lastname: Jobling
authors.forename: Jeanne
authors.initials: J
authors.lastname: Baker
authors.forename: Jennifer
authors.initials: J
authors.lastname: Johnston
authors.forename: Andrei
authors.initials: A
authors.lastname: Konradi
authors.forename: Michael P
authors.initials: MP
authors.lastname: Bova
authors.forename: Dean R
authors.initials: DR
authors.lastname: Artis
authors.forename: Rick D
authors.initials: RD
authors.lastname: Artis
mesh_terms: D004229:Dithiothreitol / Q000737:chemistry / Q000378:metabolism; D055808:Drug Discovery; D057166:High-Throughput Screening Assays / Q000379:methods; D007700:Kinetics; D008024:Ligands; D019906:Nuclear Magnetic Resonance, Biomolecular; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D019163:Reducing Agents / Q000737:chemistry / Q000378:metabolism; D020349:Surface Plasmon Resonance / Q000379:methods; D044767:Ubiquitin-Protein Ligases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism
chemical_list: D008024:Ligands; D010446:Peptide Fragments; D019163:Reducing Agents; D044767:Ubiquitin-Protein Ligases; C111567:parkin protein; D004229:Dithiothreitol