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Discovery of Small-Molecule Nonfluorescent Inhibitors of Fluorogen-Fluorogen Activating Protein Binding Pair.
Wu, Yang; Stauffer, Shaun R; Stanfield, Robyn L; Tapia, Phillip H; Ursu, Oleg; Fisher, Gregory W; Szent-Gyorgyi, Christopher; Evangelisti, Annette; Waller, Anna; Strouse, J Jacob; Carter, Mark B; Bologa, Cristian; Gouveia, Kristine; Poslusney, Mike; Waggoner, Alan S; Lindsley, Craig W; Jarvik, Jonathan W; Sklar, Larry A.
Afiliação
  • Wu Y; Department of Pathology, University of New Mexico, Albuquerque, NM, USA Center for Molecular Discovery, University of New Mexico, Albuquerque, NM, USA.
  • Stauffer SR; Vanderbilt Specialized Chemistry Center, Vanderbilt University Medical Center, Nashville, TN, USA.
  • Stanfield RL; Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA, USA.
  • Tapia PH; Center for Molecular Discovery, University of New Mexico, Albuquerque, NM, USA.
  • Ursu O; Center for Molecular Discovery, University of New Mexico, Albuquerque, NM, USA.
  • Fisher GW; Molecular Biosensor and Imaging Center, Carnegie Mellon University, Pittsburgh, PA, USA.
  • Szent-Gyorgyi C; Molecular Biosensor and Imaging Center, Carnegie Mellon University, Pittsburgh, PA, USA.
  • Evangelisti A; Center for Molecular Discovery, University of New Mexico, Albuquerque, NM, USA.
  • Waller A; Center for Molecular Discovery, University of New Mexico, Albuquerque, NM, USA.
  • Strouse JJ; Center for Molecular Discovery, University of New Mexico, Albuquerque, NM, USA.
  • Carter MB; Center for Molecular Discovery, University of New Mexico, Albuquerque, NM, USA.
  • Bologa C; Center for Molecular Discovery, University of New Mexico, Albuquerque, NM, USA.
  • Gouveia K; Center for Molecular Discovery, University of New Mexico, Albuquerque, NM, USA.
  • Poslusney M; Vanderbilt Specialized Chemistry Center, Vanderbilt University Medical Center, Nashville, TN, USA.
  • Waggoner AS; Molecular Biosensor and Imaging Center, Carnegie Mellon University, Pittsburgh, PA, USA Department of Biological Science, Carnegie Mellon University, Pittsburgh, PA, USA.
  • Lindsley CW; Vanderbilt Specialized Chemistry Center, Vanderbilt University Medical Center, Nashville, TN, USA.
  • Jarvik JW; Molecular Biosensor and Imaging Center, Carnegie Mellon University, Pittsburgh, PA, USA Department of Biological Science, Carnegie Mellon University, Pittsburgh, PA, USA.
  • Sklar LA; Department of Pathology, University of New Mexico, Albuquerque, NM, USA Center for Molecular Discovery, University of New Mexico, Albuquerque, NM, USA Lsklar@salud.unm.edu.
J Biomol Screen ; 21(1): 74-87, 2016 Jan.
Article em En | MEDLINE | ID: mdl-26442911
A new class of biosensors, fluorogen activating proteins (FAPs), has been successfully used to track receptor trafficking in live cells. Unlike the traditional fluorescent proteins (FPs), FAPs do not fluoresce unless bound to their specific small-molecule fluorogens, and thus FAP-based assays are highly sensitive. Application of the FAP-based assay for protein trafficking in high-throughput flow cytometry resulted in the discovery of a new class of compounds that interferes with the binding between fluorogens and FAP, thus blocking the fluorescence signal. These compounds are high-affinity, nonfluorescent analogs of fluorogens with little or no toxicity to the tested cells and no apparent interference with the normal function of FAP-tagged receptors. The most potent compound among these, N,4-dimethyl-N-(2-oxo-2-(4-(pyridin-2-yl)piperazin-1-yl)ethyl)benzenesulfonamide (ML342), has been investigated in detail. X-ray crystallographic analysis revealed that ML342 competes with the fluorogen, sulfonated thiazole orange coupled to diethylene glycol diamine (TO1-2p), for the same binding site on a FAP, AM2.2. Kinetic analysis shows that the FAP-fluorogen interaction is more complex than a homogeneous one-site binding process, with multiple conformational states of the fluorogen and/or the FAP, and possible dimerization of the FAP moiety involved in the process.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Ligação Proteica / Proteínas / Bibliotecas de Moléculas Pequenas Limite: Humans Idioma: En Revista: J Biomol Screen Ano de publicação: 2016 Tipo de documento: Article
Texto completo
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Ligação Proteica / Proteínas / Bibliotecas de Moléculas Pequenas Limite: Humans Idioma: En Revista: J Biomol Screen Ano de publicação: 2016 Tipo de documento: Article