fluorescence polarization

Summary

Summary: Measurement of the polarization of fluorescent light from solutions or microscopic specimens. It is used to provide information concerning molecular size, shape, and conformation, molecular anisotropy, electronic energy transfer, molecular interaction, including dye and coenzyme binding, and the antigen-antibody reaction.

Top Publications

  1. ncbi A high-throughput assay for signal transducer and activator of transcription 5b based on fluorescence polarization
    Judith Müller
    Max Planck Institute of Biochemistry, Department of Molecular Biology, and Munich Center for Integrated Protein Science CIPSM, Am Klopferspitz 18, 82152 Martinsried, Germany
    Anal Biochem 375:249-54. 2008
  2. ncbi Fluorescent protein FRET: the good, the bad and the ugly
    David W Piston
    Department of Molecular Physiology and Biophysics, Vanderbilt University, 702 Light Hall, Nashville, TN 37232 0615, USA
    Trends Biochem Sci 32:407-14. 2007
  3. ncbi Palytoxin detection and quantification using the fluorescence polarization technique
    A Alfonso
    Department of Pharmacology, Veterinary School, 27002 Lugo, Spain
    Anal Biochem 424:64-70. 2012
  4. pmc A multi-functional imaging approach to high-content protein interaction screening
    Daniel R Matthews
    Division of Cancer Studies, Randall Division of Cell and Molecular Biophysics, King s College London, London, United Kingdom
    PLoS ONE 7:e33231. 2012
  5. pmc Fluorescence polarization and fluctuation analysis monitors subunit proximity, stoichiometry, and protein complex hydrodynamics
    Tuan A Nguyen
    Section on Cellular Biophotonics, Laboratory of Molecular Physiology, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Rockville, Maryland, United States of America
    PLoS ONE 7:e38209. 2012
  6. pmc High-throughput fluorescence polarization assay to identify inhibitors of Cbl(TKB)-protein tyrosine kinase interactions
    Eric A Kumar
    Eppley Institute for Cancer Research and Allied Diseases, University of Nebraska Medical Center, Omaha, NE 68198, USA
    Anal Biochem 411:254-60. 2011
  7. ncbi Fluorescence polarization in studies of bacterial cytoplasmic membrane fluidity under environmental stress
    N C S Mykytczuk
    Department of Biology, Laurentian University, Sudbury, Ontario, Canada P3E 2C6
    Prog Biophys Mol Biol 95:60-82. 2007
  8. pmc Sensing peptide-oligonucleotide interactions by a two-color fluorescence label: application to the HIV-1 nucleocapsid protein
    Volodymyr V Shvadchak
    Laboratoire de Biophotonique et Pharmacologie, Faculte de Pharmacie, UMR 7213 du CNRS, Universite de Strasbourg, 67401 Illkirch, France
    Nucleic Acids Res 37:e25. 2009
  9. pmc Quantitative approaches to monitor protein-nucleic acid interactions using fluorescent probes
    John M Pagano
    Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA
    RNA 17:14-20. 2011
  10. pmc Analysis of protein-ligand interactions by fluorescence polarization
    Ana M Rossi
    Department of Pharmacology, University of Cambridge, Cambridge, UK
    Nat Protoc 6:365-87. 2011

Research Grants

  1. A Fluorescence Polarization Assay for Fatty Acid Adenylating Enzymes
    Courtney C Aldrich; Fiscal Year: 2012
  2. The mechanism of mRNA recruitment to the human ribosome.
    Christopher S Fraser; Fiscal Year: 2013
  3. NOVEL THERAPEUTICS FOR PRION DISEASES
    Stanley B Prusiner; Fiscal Year: 2013
  4. DNA-Binding Protein Discovery by the Combinatorial Method REPSA
    Michael van Dyke; Fiscal Year: 2013
  5. CARP-1: A Potential Therapeutic Agent for Breast Cancer
    ARUN KUMAR RISHI; Fiscal Year: 2013
  6. Homogenous HTS Assays to Screen for Inhibitors of Keap1-Nrf2 Interaction
    Longqin Hu; Fiscal Year: 2010
  7. Dual Function of VPO1 in Pathogen Recognition and Killing
    Guangjie Cheng; Fiscal Year: 2013
  8. Regulation of PKR by Novel RNA Motifs
    Philip C Bevilacqua; Fiscal Year: 2012
  9. Assays for Discovery of Translation Inhibitors
    Gerhard Wagner; Fiscal Year: 2009
  10. HTS for Rpn11 chemical probes
    Raymond J Deshaies; Fiscal Year: 2012

Detail Information

Publications331 found, 100 shown here

  1. ncbi A high-throughput assay for signal transducer and activator of transcription 5b based on fluorescence polarization
    Judith Müller
    Max Planck Institute of Biochemistry, Department of Molecular Biology, and Munich Center for Integrated Protein Science CIPSM, Am Klopferspitz 18, 82152 Martinsried, Germany
    Anal Biochem 375:249-54. 2008
    ..We have developed a high-throughput assay based on fluorescence polarization that allows screening of chemical libraries for compounds that inhibit STAT5b by interfering with the ..
  2. ncbi Fluorescent protein FRET: the good, the bad and the ugly
    David W Piston
    Department of Molecular Physiology and Biophysics, Vanderbilt University, 702 Light Hall, Nashville, TN 37232 0615, USA
    Trends Biochem Sci 32:407-14. 2007
    ..We also review the approaches that can be used to measure FRET, with particular emphasis on the potential artifacts associated with each approach...
  3. ncbi Palytoxin detection and quantification using the fluorescence polarization technique
    A Alfonso
    Department of Pharmacology, Veterinary School, 27002 Lugo, Spain
    Anal Biochem 424:64-70. 2012
    ..Its mechanism of action targets Na,K-ATPase. Fluorescence polarization (FP) is a spectroscopic technique that can be used to determine molecular interactions...
  4. pmc A multi-functional imaging approach to high-content protein interaction screening
    Daniel R Matthews
    Division of Cancer Studies, Randall Division of Cell and Molecular Biophysics, King s College London, London, United Kingdom
    PLoS ONE 7:e33231. 2012
    ..Direct correlation between acceptor anisotropy and donor FLIM is observed for FRET assays, providing an opportunity to rapidly screen proteins, interacting on the nano-meter scale, using wide-field imaging...
  5. pmc Fluorescence polarization and fluctuation analysis monitors subunit proximity, stoichiometry, and protein complex hydrodynamics
    Tuan A Nguyen
    Section on Cellular Biophotonics, Laboratory of Molecular Physiology, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Rockville, Maryland, United States of America
    PLoS ONE 7:e38209. 2012
    ..To overcome these limitations we have developed Fluorescence Polarization and Fluctuation Analysis (FPFA), a hybrid single-molecule based method combining time-resolved ..
  6. pmc High-throughput fluorescence polarization assay to identify inhibitors of Cbl(TKB)-protein tyrosine kinase interactions
    Eric A Kumar
    Eppley Institute for Cancer Research and Allied Diseases, University of Nebraska Medical Center, Omaha, NE 68198, USA
    Anal Biochem 411:254-60. 2011
    ..Here we report the development of a competitive high-throughput fluorescence polarization assay in a 384-well format to identify inhibitors of Cbl(TKB)...
  7. ncbi Fluorescence polarization in studies of bacterial cytoplasmic membrane fluidity under environmental stress
    N C S Mykytczuk
    Department of Biology, Laurentian University, Sudbury, Ontario, Canada P3E 2C6
    Prog Biophys Mol Biol 95:60-82. 2007
    ..b>Fluorescence polarization methods are valuable for measuring bacterial cytoplasmic membrane fluidity...
  8. pmc Sensing peptide-oligonucleotide interactions by a two-color fluorescence label: application to the HIV-1 nucleocapsid protein
    Volodymyr V Shvadchak
    Laboratoire de Biophotonique et Pharmacologie, Faculte de Pharmacie, UMR 7213 du CNRS, Universite de Strasbourg, 67401 Illkirch, France
    Nucleic Acids Res 37:e25. 2009
    ..The described methodology could become a new universal tool for investigating peptide-ODN interactions...
  9. pmc Quantitative approaches to monitor protein-nucleic acid interactions using fluorescent probes
    John M Pagano
    Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA
    RNA 17:14-20. 2011
    ..We review two complementary experimental methods, fluorescence polarization and fluorescence electrophoretic mobility shift assays, that enable the quantitative measurement of ..
  10. pmc Analysis of protein-ligand interactions by fluorescence polarization
    Ana M Rossi
    Department of Pharmacology, University of Cambridge, Cambridge, UK
    Nat Protoc 6:365-87. 2011
    ..b>Fluorescence polarization (FP) provides a nondisruptive means of measuring the association of a fluorescent ligand with a larger ..
  11. ncbi Development and optimization of a binding assay for the XIAP BIR3 domain using fluorescence polarization
    Zaneta Nikolovska-Coleska
    University of Michigan Comprehensive Cancer Center, Departments of Internal Medicine and Medicinal Chemistry, University of Michigan, 1500 E Medical Center Drive, Ann Arbor, MI 48109 0934, USA
    Anal Biochem 332:261-73. 2004
    ..Herein, we report the development of a homogeneous high-throughput assay based on fluorescence polarization for measuring the binding affinities of small-molecule inhibitors to the BIR3 domain of XIAP...
  12. ncbi Affinity of galectin-8 and its carbohydrate recognition domains for ligands in solution and at the cell surface
    Susanne Carlsson
    Section of Microbiology Immunology and Glycobiology MIG, Solvegatan 23, 223 62, Sweden
    Glycobiology 17:663-76. 2007
    ..More importantly, using fluorescence polarization, we also gained information on how the affinity is built by multiple weak interactions between different ..
  13. pmc Mapping the N-terminal residues of Epstein-Barr virus gp42 that bind gH/gL by using fluorescence polarization and cell-based fusion assays
    Fengling Liu
    Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305, USA
    J Virol 84:10375-85. 2010
    ..Data from fluorescence polarization, cell-cell fusion, and viral infection assays demonstrated that 33 residues corresponding to 44 to 61 ..
  14. ncbi A simple assay for hyaluronidase activity using fluorescence polarization
    Toshiyuki Murai
    Department of Immunology and Microbiology, Graduate School of Medicine, Osaka University, 2 2 Yamada oka, Suita, Osaka 565 0871, Japan
    Biochem Biophys Res Commun 376:620-4. 2008
    ..A very simple assay was established to measure degradation of hyaluronan based on fluorescence polarization. The developed assay method would provide a way for continuous measurement of cellular hyaluronidase ..
  15. pmc Homodimerization of amyloid precursor protein at the plasma membrane: a homoFRET study by time-resolved fluorescence anisotropy imaging
    Viviane Devauges
    Institut des Sciences Moléculaires d Orsay, CNRS UMR 8214, Univ Paris Sud, Orsay, France
    PLoS ONE 7:e44434. 2012
    ..This original set-up thus offers promising prospects for future studies of protein homodimerization in living cells in control and pathological conditions...
  16. pmc Homo-FRET microscopy in living cells to measure monomer-dimer transition of GFP-tagged proteins
    I Gautier
    Institut Jacques Monod, Centre National de la Recherche Scientifique, , 75251 Paris, France
    Biophys J 80:3000-8. 2001
    ..Picosecond time-resolved fluorescence anisotropy microscopy opens a promising avenue for obtaining structural information on proteins in individual living cells, even when expression levels are very low...
  17. pmc A substrate-free activity-based protein profiling screen for the discovery of selective PREPL inhibitors
    Anna Mari Lone
    Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138, USA
    J Am Chem Soc 133:11665-74. 2011
    ..Here, we use a fluorescence polarization activity-based protein profiling (fluopol-ABPP) assay to discover selective small-molecule inhibitors of ..
  18. ncbi High-throughput fluorescence polarization method for identification of FKBP12 ligands
    S Bollini
    Department of Neuroscience, Pharmaceutical Research Institute, Bristol Myers Squibb, Inc, Wallingford, CT, USA
    J Biomol Screen 7:526-30. 2002
    ..A high-throughput fluorescence polarization assay was developed to identify novel ligands of FKBP12...
  19. ncbi First direct fluorescence polarization assay for the detection and quantification of spirolides in mussel samples
    Paz Otero
    Departamento de Farmacologia, Facultad de Veterinaria, Universidad de Santiago de Compostela, Campus Universitario s n, 27002 Lugo, Spain
    Anal Chim Acta 701:200-8. 2011
    ..Other liposoluble toxins did not interfere with the assay, proving a specific method. Moreover, the matrix do not affect in the range of toxin concentrations that involving risk of spirolides intoxication...
  20. pmc ATP binding to the motor domain from an ABC transporter drives formation of a nucleotide sandwich dimer
    Paul C Smith
    Department of Biological Sciences, 702A Fairchild Center, MC2434, Columbia University, New York, New York 10027, USA
    Mol Cell 10:139-49. 2002
    ..These results establish the stereochemical basis of the power stroke of ABC transporter pumps...
  21. pmc High-throughput screening for small-molecule inhibitors of LARG-stimulated RhoA nucleotide binding via a novel fluorescence polarization assay
    CHRIS R EVELYN
    Department of Pharmacology, University of Michigan Medical Center, Ann Arbor, Michigan 48109 0632, USA
    J Biomol Screen 14:161-72. 2009
    ..Using a novel fluorescence polarization guanine nucleotide-binding assay using BODIPY-Texas Red-GTPgammaS (BODIPY-TR-GTPgammaS), the authors ..
  22. pmc A fluorescence polarization assay using an engineered human respiratory syncytial virus F protein as a direct screening platform
    Minyoung Park
    Department of Chemical and Systems Biology, Stanford University School of Medicine, CA 94305, USA
    Anal Biochem 409:195-201. 2011
    ..Here we report the development of a fluorescence polarization (FP) assay using an engineered hRSV F protein 5-helix bundle (5HB)...
  23. pmc Identification of influenza endonuclease inhibitors using a novel fluorescence polarization assay
    Brandi M Baughman
    Integrated Program in Biomedical Sciences, University of Tennessee Health Science Center, Memphis, Tennessee 38163, United States
    ACS Chem Biol 7:526-34. 2012
    ..To facilitate the discovery of endonuclease inhibitors, we have developed a high-throughput fluorescence polarization (FP) assay, utilizing a novel fluorescein-labeled compound (K(d) = 0...
  24. ncbi HomoFRET fluorescence anisotropy imaging as a tool to study molecular self-assembly in live cells
    Fiona T S Chan
    Department of Chemical Engineering and Biotechnology, University of Cambridge, Pembroke Street, Cambridge, CB2 3RA, UK
    Chemphyschem 12:500-9. 2011
    ....
  25. pmc Non-Watson-Crick interactions between PNA and DNA inhibit the ATPase activity of bacteriophage T4 Dda helicase
    Alan J Tackett
    Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, 4301 West Markham Street, Little Rock, AR 72205, USA
    Nucleic Acids Res 30:950-7. 2002
    ..However, fluorescence polarization did reveal non-sequence-specific interactions between PNA and ssDNA...
  26. ncbi Interactions between cholesterol and lipids in bilayer membranes. Role of lipid headgroup and hydrocarbon chain-backbone linkage
    S Bhattacharya
    Department of Organic Chemistry, Indian Institute of Science, Bangalore
    Biochim Biophys Acta 1467:39-53. 2000
    ....
  27. ncbi Towards multiparametric fluorescent imaging of amyloid formation: studies of a YFP model of alpha-synuclein aggregation
    Tjakko J van Ham
    Department of Genetics, University of Groningen, Groningen, The Netherlands
    J Mol Biol 395:627-42. 2010
    ..Our results validate the use of fluorescent protein chimeras of AS as representative models for studying protein aggregation and offer new opportunities for the investigation of amyloid aggregation in vivo using YFP-tagged proteins...
  28. ncbi Immobilized metal ion affinity-based fluorescence polarization (IMAP): advances in kinase screening
    J Richard Sportsman
    Molecular Devices Corp, Sunnyvale, CA, USA
    Assay Drug Dev Technol 2:205-14. 2004
    The IMAP Fluorescence Polarization technology is a homogeneous antibody-free method for analysis of kinases, phosphatases, and phosphodiesterases...
  29. pmc Characterization and use of an unprecedentedly bright and structurally non-perturbing fluorescent DNA base analogue
    Peter Sandin
    Department of Chemical and Biological Engineering Physical Chemistry, Chalmers University of Technology, SE 41296 Gothenburg, Sweden
    Nucleic Acids Res 36:157-67. 2008
    ..g. photophysical studies of DNA interacting with macromolecules (proteins) or for determining size and shape of DNA tertiary structures using techniques such as fluorescence anisotropy and fluorescence resonance energy transfer (FRET)...
  30. ncbi Probing the cathepsin D using a BODIPY FL-pepstatin A: applications in fluorescence polarization and microscopy
    C S Chen
    Molecular Probes, Inc, Eugene, OR 97402, USA
    J Biochem Biophys Methods 42:137-51. 2000
    ..The nature of its binding to cathepsin D was further characterized using a fluorescence polarization measurement. Results showed that BODIPY FL-pepstatin A selectively binds to cathepsin D at pH 4.5...
  31. ncbi The single-finger nucleocapsid protein of moloney murine leukemia virus binds and destabilizes the TAR sequences of HIV-1 but does not promote efficiently their annealing
    Caroline Egelé
    Département de Pharmacologie et Physico Chimie des Interactions Cellulaires et Moléculaires, UMR 7175 CNRS, Faculte de Pharmacie, Universite Louis Pasteur, Strasbourg 1, 74, route du Rhin, 67401 Illkirch Cedex, France
    Biochemistry 46:14650-62. 2007
    ..Due to its ability to mimic NCp7, the simple structure of NCp10 might be useful to design peptidomimetics aimed at inhibiting HIV replication...
  32. pmc Photophysical properties of Cerulean and Venus fluorescent proteins
    Pabak Sarkar
    University of North Texas Health Science Center, Center for Commercialization of Fluorescent Technologies, 3500 Camp Bowie Boulevard, Fort Worth, Texas 76107, USA
    J Biomed Opt 14:034047. 2009
    ....
  33. ncbi A high-throughput fluorescence polarization assay for signal transducer and activator of transcription 3
    Jochen Schust
    Department of Molecular Biology, Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany
    Anal Biochem 330:114-8. 2004
    ..We have developed a high-throughput binding assay, based on fluorescence polarization, which allows screening for small molecules that bind to the STAT3 SH2 domain and thereby inhibit its ..
  34. ncbi Design, synthesis, and in vitro biological evaluation of small molecule inhibitors of estrogen receptor alpha coactivator binding
    Alice L Rodriguez
    Department of Chemistry, University of Illinois, 600 South Mathews Avenue, Urbana, Illinois 61801, USA
    J Med Chem 47:600-11. 2004
    ..The pyrimidine-based CBIs appear to be the first small molecule inhibitors of NR coactivator binding...
  35. ncbi A fluorescence polarization assay for inhibitors of Hsp90
    R Howes
    Vernalis Cambridge, Granta Park, Great Abington, Cambridge CB1 6GB, UK
    Anal Biochem 350:202-13. 2006
    ..Using data from the crystal structure of an initial hit compound, we have developed a fluorescence polarization assay to monitor binding of compounds to the ATP-binding site of Hsp90...
  36. pmc Reversed-phase liquid chromatography coupled on-line to estrogen receptor bioaffinity detection based on fluorescence polarization
    Jelle Reinen
    Department of Chemistry and Pharmaceutical Sciences, LACDR Division of Molecular Toxicology, Vrije Universiteit, De Boelelaan 1083, 1081 HV, Amsterdam, The Netherlands
    Anal Bioanal Chem 390:1987-98. 2008
    ..liquid chromatography (RP-HPLC) on-line to estrogen receptor alpha (ERalpha) affinity detection using fluorescence polarization (FP)...
  37. pmc Structural changes of yellow Cameleon domains observed by quantitative FRET analysis and polarized fluorescence correlation spectroscopy
    J W Borst
    MicroSpectroscopy Centre, Laboratories of Biochemistry and Biophysics, Wageningen University, Wageningen, The Netherlands
    Biophys J 95:5399-411. 2008
    ..The combination of two independent methods gives consistent results and presents a rapid and specific methodology to analyze structural and dynamical changes in a protein on ligand binding...
  38. ncbi Fluorescence polarization and anisotropy in high throughput screening: perspectives and primer
    J C Owicki
    J Biomol Screen 5:297-306. 2000
    b>Fluorescence polarization and anisotropy are two nearly equivalent techniques that have together, over the past 5 years, achieved wide use in high throughput screening in drug discovery...
  39. pmc Identification of adiponectin receptor agonist utilizing a fluorescence polarization based high throughput assay
    Yiyi Sun
    Chengdu Medical College, Pharmacy School, Chengdu, Sichuan Province, China
    PLoS ONE 8:e63354. 2013
    ..First step, we developed a high throughput screening assay based on fluorescence polarization to identify adiponectin ligands...
  40. pmc Aryl acid adenylating enzymes involved in siderophore biosynthesis: fluorescence polarization assay, ligand specificity, and discovery of non-nucleoside inhibitors via high-throughput screening
    João Neres
    Center for Drug Design, Academic Health Center, University of Minnesota, Minneapolis, Minnesota 55455, USA
    Biochemistry 47:11735-49. 2008
    ..The probe was successfully used to develop a fluorescence polarization assay for these six AAAEs, and equilibrium dissociation constants were determined in direct binding ..
  41. ncbi Development and application of fluorescence polarization assays in drug discovery
    T J Burke
    PanVera LLC, 501 Charmany Drive, Madison, WI 53719, USA
    Comb Chem High Throughput Screen 6:183-94. 2003
    b>Fluorescence polarization technology has been used in basic research and commercial diagnostic assays for many decades, but has begun to be widely used in drug discovery only in the past six years...
  42. pmc Detection of HBV genotypes of tumor tissues and serum by a fluorescence polarization assay in north-western China's hepatocellular carcinoma patients
    Jianguo Lu
    Department of General Surgery, Tangdu Hospital, The Fourth Military Medical University, Xian, China
    Virol J 8:362. 2011
    ..The purpose of the study is to investigate hepatitis B virus genotypes distribution, the high-risk genotypes and the occult infection in north-western China's hepatocellular carcinoma patients...
  43. pmc DNA binding induces dissociation of the multimeric form of HIV-1 integrase: a time-resolved fluorescence anisotropy study
    E Deprez
    , ENS-Cachan, , 94235 Cachan, France
    Proc Natl Acad Sci U S A 98:10090-5. 2001
    ..These results indicate that upon binding the viral DNA end, the multimeric enzyme undergoes a dissociation, most likely into a homogeneous monomeric form at 25 degrees C and into a monomer-dimer equilibrium at 37 degrees C...
  44. ncbi Homogeneous cell-based fluorescence polarization assay for the direct detection of cAMP
    L Prystay
    PerkinElmer Life Sciences, Montreal, Quebec, Canada
    J Biomol Screen 6:75-82. 2001
    A fluorescence polarization-based functional assay for cyclic AMP (cAMP) production in cells has been proven effective for the detection of agonist-stimulated cAMP production in a HEK 293 recombinant cell line expressing the ..
  45. ncbi Characterization of heterotrimeric nucleotide-depleted Gα(i)-proteins by Bodipy-FL-GTPγS fluorescence anisotropy
    Lauri Tontson
    University of Tartu, Institute of Chemistry, Ravila 14a, 50411 Tartu, Estonia
    Arch Biochem Biophys 524:93-8. 2012
    ..Mn²⁺ had stabilizing effect on the nucleotide free state of the α(i1) subunit, whereas both Mn²⁺ as well as G-protein activation by mastoparan destabilized the α(i2) subunit...
  46. ncbi Fluorescence polarization assay and SDS-PAGE confirms matrilysin degrades fibronectin and collagen IV whereas gelatinase A degrades collagen IV but not fibronectin
    P J Kraft
    The R W Johnson Pharmaceutical Research Institute, Raritan, NJ 08869, USA
    Connect Tissue Res 42:149-63. 2001
    ..Changes in fluorescence intensity (FIU) and fluorescence polarization (mP) resulting from the protease activity of matrilysin and gelatinase A were measured...
  47. ncbi Development of a fluorescence polarization assay for peptidyl-tRNA hydrolase
    Paul D Bonin
    Pharmacia Corporation, Kalamazoo, Michigan 4900, USA
    Anal Biochem 306:8-16. 2002
    ..In the present study, we describe the development of a kinetic fluorescence polarization (FP) assay that enables measurements of Pth activity without the need to separate bound and free tracer...
  48. pmc A new class of rhomboid protease inhibitors discovered by activity-based fluorescence polarization
    Eliane V Wolf
    Lehrstuhl für Chemie der Biopolymere, Technische Universitat Munchen, Freising, Germany
    PLoS ONE 8:e72307. 2013
    ..To overcome the need for substrates, we developed a screening assay using fluorescence polarization activity-based protein profiling (FluoPol ABPP) that is compatible with membrane proteases...
  49. pmc Steady-state polarization from cylindrically symmetric fluorophores undergoing rapid restricted motion
    M Irving
    Randall Institute, King s College London, England
    Biophys J 70:1830-5. 1996
    Steady-state fluorescence polarization measurements provide a relatively simple method for investigating the orientation of molecular components in ordered biological systems...
  50. ncbi Single-stranded DNA binding protein-assisted fluorescence polarization aptamer assay for detection of small molecules
    Zhenyu Zhu
    Département de Pharmacochimie Moléculaire UMR 5063, Institut de Chimie Moleculaire de Grenoble FR 2607, CNRS Universite Grenoble I Joseph Fourier, 38041 Grenoble Cedex 9, France
    Anal Chem 84:7203-11. 2012
    Here, we describe a new fluorescence polarization aptamer assay (FPAA) strategy which is based on the use of the single-stranded DNA binding (SSB) protein from Escherichia coli as a strong FP signal enhancer tool...
  51. pmc Comprehensive binary interaction mapping of SH2 domains via fluorescence polarization reveals novel functional diversification of ErbB receptors
    Ronald J Hause
    the Committee on genetics, Genomics, and Systems Biology, The Ben May Department for Cancer Research and the Institute for Genomics and Systems Biology, The Gwen and Jules Knapp Center for Biomedical Discovery, The University of Chicago, Chicago, Illinois, United States of America
    PLoS ONE 7:e44471. 2012
    ..Here, we developed a large-scale fluorescence polarization (FP) methodology that was able to characterize interactions between SH2 domains and ErbB receptor ..
  52. ncbi Asymmetric GP5+/6+ PCR and hybridization with fluorescence polarization assay of 15 human papillomavirus genotypes in clinical samples
    Zhang Ju
    Institute of Gene Diagnosis, State Key Laboratory of Cancer Biology, Fourth Military Medical University, 17 Changle West Road, Xian, Shaanxi 710032, China
    J Clin Virol 44:106-10. 2009
    ..The detection of a broad spectrum of HPV genotypes has not been used widely in cervical cancer screening due to technical difficulties and high costs...
  53. ncbi Application of fluorescence polarization to the steady-state enzyme kinetic analysis of calpain II
    D S Sem
    La Jolla Pharmaceutical Company, San Diego, CA 92121, USA
    FEBS Lett 443:17-9. 1999
    This paper presents the application of fluorescence polarization to the determination of dissociation constants for competitive inhibitors that bind to enzymes...
  54. ncbi Fluorescence polarization is a useful technology for reagent reduction in assay miniaturization
    T J Kowski
    Novartis Institute for Biomedical Research, Novartis Pharmaceuticals Corporation, Summit, NJ 07901, USA
    Comb Chem High Throughput Screen 3:437-44. 2000
    The use of fluorescence polarization (FP) has increased significantly in the development of sensitive and robust assays for high throughput screening of chemical compound libraries during the past few years...
  55. ncbi Detection of 13,19-didesmethyl C spirolide by fluorescence polarization using Torpedo electrocyte membranes
    Eva S Fonfría
    Departamento de Farmacologia, Facultad de Veterinaria, Universidad de Santiago de Compostela, Campus Universitario, 27002 Lugo, Spain
    Anal Biochem 403:102-7. 2010
    b>Fluorescence polarization (FP) is a powerful tool for studying molecular interactions by monitoring changes in the apparent size of fluorescent molecules...
  56. pmc Fluorescence polarization in homogeneous nucleic acid analysis II: 5'-nuclease assay
    S Latif
    Division of Dermatology, Washington University School of Medicine, St. Louis, Missouri 63110, USA
    Genome Res 11:436-40. 2001
    When the temperature and viscosity of the solvent is held constant, the degree of fluorescence polarization (FP) detected when a fluorescent dye is excited by plane polarized light depends mostly on the molecular weight of the dye ..
  57. ncbi Fluorescence anisotropy assay for pharmacological characterization of ligand binding dynamics to melanocortin 4 receptors
    Santa Veiksina
    Department of Bioorganic Chemistry, Institute of Chemistry, University of Tartu, 50411 Tartu, Estonia
    Anal Biochem 402:32-9. 2010
    ..Real-time monitoring of ligand binding dynamics enabled us to find optimal experimental conditions for each particular ligand and an improved estimate of their binding parameters...
  58. ncbi Fluorescence polarization analysis for revealing molecular mechanism of nucleotide-dependent phospholipid membrane binding of MinD adenosine 5'-triphosphate, adenosine triphosphatase
    Takashi Okuno
    Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Japan
    Biol Pharm Bull 33:1746-50. 2010
    ..Our results presumed that K11 and K16 residues might play an important role in dimmer formation of MinD, but not ATP binding step, for recruiting to membrane...
  59. ncbi Improved photostable FRET-competent biarsenical-tetracysteine probes based on fluorinated fluoresceins
    Carla C Spagnuolo
    Departamento de Quimica Organica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, CIHIDECAR, CONICET, Ciudad Universitaria Pabellon II, 1428, Buenos Aires, Argentina
    J Am Chem Soc 128:12040-1. 2006
    ..The two probes also provide a new FRET pair with a larger Ro value (54 A) than any previously obtained with biarsenical dyes...
  60. ncbi Effects of oncogenic mutations and DNA response elements on the binding of p53 to p53-binding protein 2 (53BP2)
    Henning Tidow
    Centre for Protein Engineering, Medical Research Council, Hills Road, Cambridge CB2 2QH, United Kingdom
    J Biol Chem 281:32526-33. 2006
    ..We found that binding of 53BP2 and DNA to p53 is mutually exclusive in the case of GADD45, p21, Bax, and PIG3. Both pro-apoptotic and non-apoptotic response elements were competed off p53 by 53BP2 with no indication of a ternary complex...
  61. ncbi Time-lapse FRET microscopy using fluorescence anisotropy
    D R Matthews
    Richard Dimbleby Department of Cancer Research, New Hunts House, Kings College London, Guy s Medical School Campus, SE11UL, UK
    J Microsc 237:51-62. 2010
    ....
  62. pmc Use of recombinant flagellin protein as a tracer antigen in a fluorescence polarization assay for diagnosis of leptospirosis
    N I Bughio
    Canadian Food Inspection Agency, Animal Diseases Research Institute, Nepean, Ontario, Canada K2H 8P9
    Clin Diagn Lab Immunol 6:599-605. 1999
    ..protein, FlaB, of Leptospira interrogans serovar pomona in the serodiagnosis of leptospirosis by the fluorescence polarization assay (FPA)...
  63. ncbi Development of a high-throughput fluorescence polarization assay for the discovery of phosphopantetheinyl transferase inhibitors
    Benjamin P Duckworth
    Center for Drug Design, University of Minnesota, Minneapolis, MN 55455, USA
    Anal Biochem 403:13-9. 2010
    ..Here we describe the development of a fluorescence polarization (FP) assay that is amenable for high-throughput screening to identify PPTase inhibitors...
  64. ncbi Protein-binding aptamer assisted signal amplification for the detection of influenza A (H1N1) DNA sequences based on quantum dot fluorescence polarization analysis
    Juanni Zhang
    Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources Ministry of Education of China, College of Chemistry and Chemical Engineering of Guangxi Normal University, Guilin, 541004, China
    Analyst 138:4722-7. 2013
    We report a fluorescence polarization platform for H1N1 detection based on the construction of a DNA functional QD fluorescence polarization probe and a bi-functional protein binding aptamer (Apt-DNA)...
  65. ncbi Optimization of the structure-switching aptamer-based fluorescence polarization assay for the sensitive tyrosinamide sensing
    Zhenyu Zhu
    Département de Pharmacochimie Moléculaire UMR 5063 CNRS, ICMG FR 2607, Universite Grenoble I, Saint Martin d Heres, France
    Anal Chim Acta 707:191-6. 2011
    In this paper, a structure-switching aptamer assay based on a fluorescence polarization (FP) signal transduction approach and dedicated to the L-tyrosinamide sensing was described and optimized...
  66. ncbi Competitive fluorescence polarization assays for the detection of phosphoinositide kinase and phosphatase activity
    Beth E Drees
    Echelon Biosciences, Inc, 675 Arapeen Drive, Salt Lake City, UT 84108, USA
    Comb Chem High Throughput Screen 6:321-30. 2003
    We describe the development and implementation of competitive fluorescence polarization (FP) based assays for determining activity of phosphoinositide 3-kinase (PI 3-K) and the type-II SH2-domain-containing inositol 5-phosphatase (SHIP2)...
  67. ncbi A TR3/Nur77 peptide-based high-throughput fluorescence polarization screen for small molecule Bcl-B inhibitors
    Kenneth W Yip
    Burnham Institute for Medical Research, La Jolla, California 92037, USA
    J Biomol Screen 13:665-73. 2008
    ..TR3-r8 peptide was used to screen for small molecule Bcl-B inhibitors. A fluorescence polarization assay (FPA) employing fluorescein isothiocyanate (FITC)-labeled TR3-r8 peptide (FITC-TR3-r8) and Bcl-B ..
  68. pmc A high-throughput fluorescence polarization anisotropy assay for the 70N domain of replication protein A
    Elaine M Souza-Fagundes
    Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN 37232, USA
    Anal Biochem 421:742-9. 2012
    ..Here we report on the development of a homogeneous, high-throughput fluorescence polarization assay for identifying compounds that block the critical protein-protein interaction site in the basic ..
  69. ncbi A general framework for development and data analysis of competitive high-throughput screens for small-molecule inhibitors of protein-protein interactions by fluorescence polarization
    Michael H A Roehrl
    Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA
    Biochemistry 43:16056-66. 2004
    ..We discuss a general framework for high-throughput screening assays based on fluorescence polarization and strategies for assay development, sensitivity regimes, data quality control, analysis, and ranking...
  70. ncbi Effects of chlorpromazine on plasma membrane permeability and fluidity in the rat brain: a dynamic positron autoradiography and fluorescence polarization study
    Nobuyuki Maruoka
    Department of Neuropsychiatry, University of Fukui, Fukui 910 1193, Japan
    Prog Neuropsychopharmacol Biol Psychiatry 31:178-86. 2007
    ..CPZ loading dose-dependently increased both membrane permeability and membrane fluidity, which suggested the involvement of a perturbation of membrane order in the mechanisms of membrane destabilization induced by antipsychotic drugs...
  71. ncbi Measurement of protease activity of live Uronema marinun (Ciliata: Scuticociliatida) by fluorescence polarization
    Eun Hye Lee
    Department of Aquatic Life Medicine, Pukyong National University, Pusan 808 737, Korea
    Dis Aquat Organ 54:85-8. 2003
    The proteolytic activity of live Uronema marinum was analyzed by a fluorescence polarization (FP) technique...
  72. ncbi Analysis of ligand-dependent recruitment of coactivator peptides to estrogen receptor using fluorescence polarization
    Mary Szatkowski Ozers
    Invitrogen Corporation, 501 Charmany Drive, Madison, Wisconsin 53719, USA
    Mol Endocrinol 19:25-34. 2005
    ..random phage display sequences were used with purified ERalpha or ERbeta in an in vitro high-throughput fluorescence polarization assay...
  73. ncbi Stress-induced alteration of chlorophyll fluorescence polarization and spectrum in leaves of Alocasia macrorrhiza L. Schott
    Zhi Fang Lin
    South China Botanical Garden, Chinese Academy of Sciences, Guangdong Key Laboratory of Digital Garden, Guangzhou, China
    J Fluoresc 17:663-9. 2007
    The value of intrinsic chlorophyll fluorescence polarization, and the intensity in emission spectrum were investigated in leaf segments of Alocasia macrorrhiza under several stress conditions including different temperatures (25-50 ..
  74. pmc PDZ domain binding selectivity is optimized across the mouse proteome
    Michael A Stiffler
    Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, MA 02138, USA
    Science 317:364-9. 2007
    ..We used protein microarrays and quantitative fluorescence polarization to characterize the binding selectivity of 157 mouse PDZ domains with respect to 217 genome-encoded ..
  75. pmc Simultaneously monitoring DNA binding and helicase-catalyzed DNA unwinding by fluorescence polarization
    H Q Xu
    Laboratoire de Biotechnologies et Pharmacologie génétique Appliquée, CNRS UMR 8532, Ecole Normale Supérieure ENS de Cachan, 61 Avenue du President Wilson, 94235 Cachan Cedex, France
    Nucleic Acids Res 31:e70. 2003
    ..This assay takes advantage of the substantial change in fluorescence polarization (FP) upon helicase binding and DNA unwinding...
  76. ncbi Use of fluorescence polarization to monitor MHC-peptide interactions in solution
    S Dedier
    Department of Applied BioSciences, Swiss Federal Institute of Technology, Winterthurerstrasse 190, CH 8057, Zurich, Switzerland
    J Immunol Methods 255:57-66. 2001
    We describe here fluorescence polarization-based methods to investigate class I MHC-peptide interactions in solution...
  77. ncbi Influence of MT7 toxin on the oligomerization state of the M1 muscarinic receptor
    Catherine Marquer
    CEA, iBiTec S, Service d Ingenierie Moleculaire des Proteines SIMOPRO, Gif sur Yvette, F 91191, France
    Biol Cell 102:409-20. 2010
    ..We chose to focus our present study on the effect of MT7 (muscarinic toxin 7), a highly selective allosteric peptide ligand, on the oligomerization state of the hM1 (human M1 muscarinic acetylcholine receptor subtype)...
  78. ncbi Structural characterization of liposomes made of diether archaeal lipids and dipalmitoyl-L-α-phosphatidylcholine
    Dejan Gmajner
    University of Ljubljana, Slovenia
    Biophys Chem 158:150-6. 2011
    ..Photon correlation spectroscopy and transmission electron microscopy show that mixed liposomes do not fuse or aggregate, even after 41 days at 4 °C...
  79. pmc Binding of inositol 1,4,5-trisphosphate (IP3) and adenophostin A to the N-terminal region of the IP3 receptor: thermodynamic analysis using fluorescence polarization with a novel IP3 receptor ligand
    Zhao Ding
    Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge, CB2 1PD, United Kingdom
    Mol Pharmacol 77:995-1004. 2010
    ..We therefore developed a high-throughput fluorescence polarization (FP) assay using a newly synthesized analog of IP(3), fluorescein isothiocyanate (FITC)-IP(3), to ..
  80. ncbi Application of fluorescence polarization in HTS assays
    Xinyi Huang
    Chemical and Screening Sciences, Wyeth Research, Collegeville, PA, USA
    Methods Mol Biol 565:127-43. 2009
    Steady-state measurements of fluorescence polarization have been widely adopted in the field of high-throughput screening for the study of biomolecular interactions...
  81. ncbi Displacement of protein-bound aptamers with small molecules screened by fluorescence polarization
    Markus Hafner
    Program Unit Chemical Biology and Medicinal Chemistry, LIMES Institute, c o Kekulé Institut für Organische Chemie und Biochemie, University of Bonn, Gerhard Domagk Strasse 1, 53121 Bonn, Germany
    Nat Protoc 3:579-87. 2008
    ..The method relies on fluorescence-labeled aptamers and the increase of fluorescence polarization upon their binding to the target protein...
  82. pmc Probing the dynamic organization of transcription compartments and gene loci within the nucleus of living cells
    Deepak Kumar Sinha
    National Centre for Biological Sciences, Tata Institute of Fundamental Research, Bangalore, India
    Biophys J 95:5432-8. 2008
    ..This heterogeneous behavior in TC dynamics within living cells could provide an interesting paradigm to explore the spatiotemporal dimension to gene transcription control...
  83. pmc A high throughput fluorescence polarization assay for inhibitors of the GoLoco motif/G-alpha interaction
    Adam J Kimple
    NIH Chemical Genomics Center, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892 3370, USA
    Comb Chem High Throughput Screen 11:396-409. 2008
    ..Here, we describe a high throughput fluorescence polarization (FP) assay using fluorophore-labeled GoLoco motif peptides for identifying inhibitors of the GoLoco ..
  84. ncbi Heterogeneous transition metal-based fluorescence polarization (HTFP) assay for probing protein interactions
    Alexander Riechers
    Institute for Organic Chemistry, University of Regensburg, Regensburg, Germany
    Biotechniques 47:837-44. 2009
    ..The present study details the development of a fluorescence polarization assay using melanoma inhibitory activity (MIA) protein-binding compounds and studies of the binding ..
  85. pmc Fluorescence polarization method to characterize macrolide-ribosome interactions
    Kang Yan
    1250 South Collegeville Road, Collegeville, PA 19426, USA
    Antimicrob Agents Chemother 49:3367-72. 2005
    A fluorescence polarization assay is described that measures the binding of fluorescently labeled erythromycin to 70S ribosomes from Escherichia coli and the displacement of the erythromycin from these ribosomes...
  86. ncbi Real-time measurement of in vitro peptide binding to soluble HLA-A*0201 by fluorescence polarization
    Rico Buchli
    Pure Protein L L C, Oklahoma City, Oklahoma 73104 3698, USA
    Biochemistry 43:14852-63. 2004
    ..Here, we report the development of a sensitive biochemical assay that relies upon fluorescence polarization to indicate peptide interactions with recombinant soluble HLA proteins...
  87. ncbi A high-throughput screen for identification of molecular mimics of Smac/DIABLO utilizing a fluorescence polarization assay
    Constance J Glover
    Developmental Therapeutics Program, National Cancer Institute at Frederick, Frederick, MD 21702, USA
    Anal Biochem 320:157-69. 2003
    ..We have developed a high-throughput fluorescence polarization assay utilizing a fluorescein-labeled peptide similar to the "IAP binding" domain of Smac N terminus ..
  88. ncbi Detection of hepatitis B virus genotypes A to D by the fluorescence polarization assay based on asymmetric PCR
    Li Ding
    The Center of Biotechnological Diagnosis and Therapy, The 261st Hospital of PLA, Beijing, China
    J Virol Methods 168:31-7. 2010
    ..fluorophores hybridized, respectively with their target amplicons, and the hybridization increased the fluorescence polarization (FP) values. The genotypes were determined by the increased fluorescence polarization values...
  89. ncbi Evaluation of the fluorescence polarization assay and comparison to other serological assays for detection of brucellosis in cervids
    D Gall
    Canadian Food Inspection Agency, Animal Diseases Research Institute, Nepean, Ontario
    J Wildl Dis 37:110-8. 2001
    ..fixation test (CFT), competitive enzyme immunoassay (CELISA), indirect enzyme immunoassay (IELISA) and fluorescence polarization assay (FPA) were evaluated for the detection of antibodies to Brucella abortus and Brucella suis biotype ..
  90. ncbi Rapid and high throughput detection of HBV YMDD mutants with fluorescence polarization
    Yui Jie Bai
    Institute of Genetic Diagnosis, Fourth Military Medical University, Xi an 710032, Shannxi Province, China
    World J Gastroenterol 9:2344-7. 2003
    ..To develop a simple and rapid detection of HBV gene variants and prediction of lamivudine-resistance in patients...
  91. ncbi Development of a fluorescence polarization assay to screen for inhibitors of the FtsZ/ZipA interaction
    Cynthia Hess Kenny
    Screening Sciences, Biophysics Enzymology, Wyeth Research, Pearl River, NY 10965, USA
    Anal Biochem 323:224-33. 2003
    A fluorescence polarization competition assay has been developed to screen for inhibitors of the Escherichia coli FtsZ/ZipA protein-protein interaction...
  92. ncbi Determination of drug-serum protein interactions via fluorescence polarization measurements
    Ulf Mathias
    Institute of Pharmaceutical Sciences, University of Freiburg, Albertstr 25, Freiburg, Germany
    Anal Bioanal Chem 388:1147-56. 2007
    ..It is based on selective fluorescent probes and the measurement of fluorescence polarization. This leads to decreased interference with fluorescent drugs as compared with previously published ..
  93. ncbi Efficient and expedient two-step pyranose-retaining fluorescein conjugation of complex reducing oligosaccharides: galectin oligosaccharide specificity studies in a fluorescence polarization assay
    Christopher T Oberg
    Department of Chemistry, University of Waterloo, 200 University Avenue West, Waterloo, Ontario N2L 3G1, Canada
    Bioconjug Chem 14:1289-97. 2003
    ..run against galectins-1, -3, and -8, beta-galactoside recognizing lectins of current interest, in a fluorescence polarization binding assay...
  94. ncbi A fluorescence polarization assay for the identification of inhibitors of the p53-DM2 protein-protein interaction
    Stephen M G Knight
    Department of Pharmacology, University of Wisconsin Madison, Madison, Wisconsin 53706 1532, USA
    Anal Biochem 300:230-6. 2002
    ..To assist this effort, we have devised a fluorescence polarization assay that quantifies the interaction between the N-terminal regions of both proteins in 384-well ..
  95. ncbi FRET by fluorescence polarization microscopy
    David W Piston
    Department of Molecular Physiology and Biophysics, Vanderbilt University Medical Center, Nashville, Tennessee 37232, USA
    Methods Cell Biol 85:415-30. 2008
    ..In this chapter, we will discuss the theory behind AFRET imaging, calculation of FP anisotropies using fluorescent microscopes, and configuration of microscopes for AFRET experiments...
  96. pmc Single nucleotide polymorphism genotyping using short, fluorescently labeled locked nucleic acid (LNA) probes and fluorescence polarization detection
    Anton Simeonov
    Caliper Technologies Corporation, Mountain View, CA 94043, USA
    Nucleic Acids Res 30:e91. 2002
    ..or hexachlorofluorescein (HEX), and their hybridization to target DNAs is followed by measuring the fluorescence polarization (FP) of the dyes...
  97. pmc Adaptability of myosin V studied by simultaneous detection of position and orientation
    Sheyum Syed
    Department of Physics, University of Illinois, Urbana Champaign, 61801, USA
    EMBO J 25:1795-803. 2006
    ....
  98. ncbi New biarsenical ligands and tetracysteine motifs for protein labeling in vitro and in vivo: synthesis and biological applications
    Stephen R Adams
    Department of Pharmacology, Department of Chemistry and Biochemistry, Howard Hughes Medical Institute, and Biomedical Sciences Program, University of California, San Diego, La Jolla, California 92093 0647, USA
    J Am Chem Soc 124:6063-76. 2002
    ..Analogous biarsenicals enable affinity chromatography, fluorescence anisotropy measurements, and electron-microscopic localization of tetracysteine-tagged proteins...
  99. ncbi Three-dimensional structural dynamics of myosin V by single-molecule fluorescence polarization
    Joseph N Forkey
    Pennsylvania Muscle Institute, University of Pennsylvania, Philadelphia, Pennsylvania 19104 6083, USA
    Nature 422:399-404. 2003
    ..of the structural dynamics of the light chain domain of brain myosin V using a single-molecule fluorescence polarization technique that determines the orientation of individual protein domains with 20-40-ms time resolution...
  100. pmc Inhibiting HIV-1 integrase by shifting its oligomerization equilibrium
    Zvi Hayouka
    Department of Organic Chemistry, Department of Biological Chemistry, Protein Purification Unit, Hebrew University of Jerusalem, Givat Ram, Jerusalem 91904, Israel
    Proc Natl Acad Sci U S A 104:8316-21. 2007
    ..These peptides are promising anti-HIV lead compounds that modulate oligomerization of IN via a previously uncharacterized mechanism, which bears advantages over the conventional interface dimerization inhibitors...
  101. pmc Uncovering quantitative protein interaction networks for mouse PDZ domains using protein microarrays
    Michael A Stiffler
    Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138, USA
    J Am Chem Soc 128:5913-22. 2006
    ..we define a gold standard dataset by determining the K(D) for all 520 PDZ-peptide combinations using fluorescence polarization. We then show that microarrays of PDZ domains identify interactions of moderate to high affinity (K(D) &..

Research Grants65

  1. A Fluorescence Polarization Assay for Fatty Acid Adenylating Enzymes
    Courtney C Aldrich; Fiscal Year: 2012
    ..Toward this goal we propose to develop a fluorescence polarization assay amenable to high throughput screening to identify small molecule FadD inhibitors...
  2. The mechanism of mRNA recruitment to the human ribosome.
    Christopher S Fraser; Fiscal Year: 2013
    ..Important amino acids will be identified using the fluorescence polarization assay we have established together with site-directed mutagenesis of eIF4G...
  3. NOVEL THERAPEUTICS FOR PRION DISEASES
    Stanley B Prusiner; Fiscal Year: 2013
    ..using HuPrPSc to seed the polymerization of recombinant (rec) HuPrP as well as a humanized version of a fluorescence polarization (FP) assay that measures the binding of PrP peptides to HuPrPSc...
  4. DNA-Binding Protein Discovery by the Combinatorial Method REPSA
    Michael van Dyke; Fiscal Year: 2013
    ..and determine binding specificity using recombinant proteins, point-mutated oligonucleotides, and fluorescence polarization binding assays...
  5. CARP-1: A Potential Therapeutic Agent for Breast Cancer
    ARUN KUMAR RISHI; Fiscal Year: 2013
    ..Following mapping of the interacting epitopes of CARP-1 and APC-2, we developed a fluorescence polarization assay (FPA) and screened a chemical library to identify small molecule inhibitors (SMIs) of CARP-1:APC-2 ..
  6. Homogenous HTS Assays to Screen for Inhibitors of Keap1-Nrf2 Interaction
    Longqin Hu; Fiscal Year: 2010
    ..high throughput screening assays will be developed and validated in this new R01 application using fluorescence polarization (FP) and time-resolved fluorescence resonance energy transfer (TR-FRET)...
  7. Dual Function of VPO1 in Pathogen Recognition and Killing
    Guangjie Cheng; Fiscal Year: 2013
    ..major methods for addressing the aims include molecular cloning and expressing, protein purification, fluorescence polarization technology and surface plasmon resonance technology...
  8. Regulation of PKR by Novel RNA Motifs
    Philip C Bevilacqua; Fiscal Year: 2012
    ..Binding assays will be conducted by fluorescence polarization and ITC;protein dimerization will be monitored by pull-down assays, crosslinking, and analytical ..
  9. Assays for Discovery of Translation Inhibitors
    Gerhard Wagner; Fiscal Year: 2009
    ..for use as a secondary screen based on directly monitoring the binding between eIF4G/eIF4A/RNA using a fluorescence polarization (FP) or scintillation proximity assays (SPA)...
  10. HTS for Rpn11 chemical probes
    Raymond J Deshaies; Fiscal Year: 2012
    ..By measuring the fluorescence polarization of the substrate in the presence of 26S proteasome plus various compounds, we should be able to identify ..
  11. Structural basis of mRNA decapping by Dcp2: conformational changes &co-activator
    JEFFREY SCOTT MUGRIDGE; Fiscal Year: 2013
    ..or metal fluoride additives will be identified using a combination of small angle x-ray scattering, fluorescence polarization and enzyme inhibition experiments...
  12. Methyltransferase inhibition assay development
    Bernard Weisblum; Fiscal Year: 2010
    ..of a fluorescent "surrogate" peptide aptamer with the target methyltransferase will be measured using fluorescence polarization. The proposed assay is a "mix- and-measure" type of determination that is based on the principle that ..
  13. Identification of inhibitors of hedgehog autoprocessing
    HENRY contact PAULUS; Fiscal Year: 2010
    ..This approach is based on an in vitro homogeneous assay system which measures changes in fluorescence polarization that accompany the cholesterol-depended autocleavage of Hh protein...
  14. Assay Development for Substrate and Phosphorylation State Specific JNK Inhibitors
    Philip LoGrasso; Fiscal Year: 2013
    ..To accomplish this we will: 1) Develop a fluorescence polarization (FP) assay for discovering substrate and bidentate competitive selective inhibitors, and fluorescence ..
  15. The Role of N-myristylation and N-terminal Modifications on PKA Structure and Dyn
    ADAM CHRISTOPHER BASTIDAS; Fiscal Year: 2013
    ..we will fluorescently label PKA with fluorescein maleimide to understand protein dynamics using fluorescence polarization. We hypothesize that these techniques will reveal new and important isoform specific dynamics of ..
  16. Small-molecule Inhibition of the Interactions of the Urokinase Receptor: A Targe
    Samy Meroueh; Fiscal Year: 2012
    ..chemical synthesis of the most promising compounds, followed by an assessment of their activity using a fluorescence polarization assay. The most potent compounds are further characterized with isothermal titration calorimetry...
  17. Molecular motors in cell biology
    Yale E Goldman; Fiscal Year: 2013
    ..Single-molecule fluorescence polarization, nanometer-resolved fluorophore localization, infrared optical traps, rapid biochemical reaction ..
  18. Apolipoprotein E/Lipoprotein Binding Mechanism
    Vasanthy Narayanaswami; Fiscal Year: 2013
    ..this, we will: (i) determine the unfolding pattern of recombinant apoE3 and apoE4 using a combination of fluorescence polarization (FP) and hydrogen deuterium exchange coupled to mass spectrometry (HDX-MS);(ii) compare the lipid ..
  19. Surface Plasmon Resonance (SPR) Biosensor- Biacore T200
    Yousef Al-Abed; Fiscal Year: 2012
    ..binding proteins for subsequent identification and characterization, extending techniques such as ELISA, fluorescence polarization, and yeast two-hybrid systems...
  20. Inhibitors of Kaposi???s Sarcoma Herpesvirus
    Kenneth M Kaye; Fiscal Year: 2011
    ..This work will develop a robust, reproducible, and miniaturized fluorescence polarization (FP) assay for a high throughput screen for small molecules that inhibit KSHV LANA binding to its DNA ..
  21. The PIP2-virus interface and PI 4-kinase: novel biology and validation targets
    Jeffrey S Glenn; Fiscal Year: 2013
    ..domain peptides to define their key determinants of PIP2 binding using quartz crystal microbalance, fluorescence polarization, and the nuclear magnetic resonance structures of the peptides;c) developing an algorithm to ..
  22. DnaK Protein Inhibitor Antimicrobials
    Michael Sturgess; Fiscal Year: 2003
    ..Agrobacterium tumefaciens DnaK by using spectroscopic and molecular modeling techniques as well as by fluorescence polarization studies between the peptides and mutated DnaK fragments...
  23. Treating type 2 diabetes with menin inhibitors
    Frank Leu; Fiscal Year: 2013
    ..These compounds will represent a new class of anti-diabetic drugs, orally administered, negating the need for repeated injections of exogenous insulin in the 17 million US patients diagnosed with T2D. ..
  24. High Throughput Assay Development for ARFGAP Modulators
    Qisheng Zhang; Fiscal Year: 2011
    ..In our efforts to dissect ARFGAP-regulated cell signaling, a novel fluorescence polarization-based ARFGAP assay has been developed. The Z'factor of the assay is 0.75 in 384-well format...
  25. High Throughput Screen For HIV-1 MA/CA Processing Site Inhibitors
    RONALD I SWANSTROM; Fiscal Year: 2011
    ..In this application we describe a fluorescence polarization assay that can monitor cleavage at the MA/CA processing site in the context of the folded protein ..
  26. Chemical Probes of Kaposi's Sarcoma Herpesvirus Latent Infection
    Kenneth M Kaye; Fiscal Year: 2010
    ..This work will develop a robust, reproducible, and miniaturized fluorescence polarization (FP) assay for a high throughput screen for small molecules that inhibit KSHV LANA binding to histones ..
  27. Reactivating Apoptosis in Refractory Cancer by Selective Targeting of MCL-1
    Michelle Stewart; Fiscal Year: 2010
    ..and dissociate its heterodimeric interactions with pro-apoptotic BCL-2 proteins such as BAK, competitive fluorescence polarization binding assays and in situ co-immunoprecipitation studies will be performed...
  28. Mechanism of Eukaryotic Environmental Mutagenesis
    Graham C Walker; Fiscal Year: 2013
    ..details of Rev1/Pol ?-dependent TLS, while at the same time allowing us to develop high-throughput fluorescence polarization assays for screening for small molecules that disrupt Rev1-Rev7-Rev3 interactions...
  29. HIVRAD: Structural approaches to vaccine development
    Ellis Reinherz; Fiscal Year: 2009
    ..Wagner will also employ fluorescence polarization assays to identify HIV gpl40-binding organic inhibitors of the CD4-gpl40 interaction...
  30. Gyrase B Inhibitors for Mycobacterium Tuberculosis
    Peter B Madrid; Fiscal Year: 2013
    ..synthesize and purify this targeted library, which will then be screened for binding to gyrase B using a fluorescence polarization binding assay...
  31. Measuring and Predicting PDZ Domain-Peptide Interactions on a Genome-wide Scale
    Gavin MacBeath; Fiscal Year: 2013
    ..In Aim 1, a strategy is presented that uses protein microarrays and high-throughput fluorescence polarization to train a statistical model that predicts PDZ domain-peptide interactions on a proteome-wide scale...
  32. Screening for Inhibitors for ERG in Prostate Cancer
    JOHN HACKETT BUSHWELLER; Fiscal Year: 2010
    ..We will use a well-validated fluorescence polarization assay to screen the MLPCN library of compounds...
  33. Mechanism of Eukaryotic Environmental Mutagenesis
    Graham C Walker; Fiscal Year: 2011
    ..of Rev1/Pol ζ-dependent TLS, while at the same time allowing us to develop high-throughput fluorescence polarization assays for screening for small molecules that disrupt Rev1-Rev7-Rev3 interactions...
  34. An Activity-Based Assay to Screen for PRMT1 Inhibitors
    Kerri A Mowen; Fiscal Year: 2013
    ..labeling with fluorescently labeled maleimide probe in the presence of inhibitors by monitoring the fluorescence polarization signal...
  35. Towards In Situ Optical Pathology of Nonmelanoma Skin Cancers
    Anna N Yaroslavsky; Fiscal Year: 2010
    ..Both modes will acquire images in reflectance, fluorescence, and fluorescence polarization. To enhance the contrast of the acquired images, we will use an FDA-approved conventional histological ..
  36. Automated high-throughput analysis system
    Satchidananda Panda; Fiscal Year: 2010
    ..The multimode detector plate reader will measure absorbance, fluorescence intensity, luminescence, fluorescence polarization and time-resolved fluorescence...
  37. Design of Macrocyclic Inhibitors of the NEMO/IKKa/b Protein-Protein Interaction
    Adrian Whitty; Fiscal Year: 2013
    ..macrocycles and acyclic inhibitors (from Aim 1), and test them for inhibition of NEMO/IKK binding in a fluorescence polarization assay...
  38. Investigation of the catalytic and non-catalytic roles of Src family kinases in o
    Stephen Leonard; Fiscal Year: 2013
    ..First, fluorescence polarization, limited proteolysis and pull-down assays will be used to probe the accessibility of the SH2 and SH3 ..
  39. Allosteric Inhibitor Discovery Technology for Protein Kinases
    THOMAS EDWARD SMITHGALL; Fiscal Year: 2013
    ..in a structurally competent state, providing an excellent starting point for novel single- molecule fluorescence polarization and/or FRET HTS assay development...
  40. Specific Chemical Probes for Histone Demethylases
    Xiang Wang; Fiscal Year: 2013
    ..The fluorescent probe is being applied to develop a fluorescence polarization-based assay, which will not only enable us to validate our bivalent inhibitor design, but also provide a ..
  41. High-throughput screening for HIV assembly and maturation inhibitors
    Wuyuan Lu; Fiscal Year: 2013
    ..seeks to fill an important technological gap in anti-HIV drug discovery by developing a sensitive fluorescence polarization assay that can be automated for high throughput screening (HTS) for small molecule inhibitors of ..
  42. The Electrophysiological Studies of Voltage Gated Channels
    FRANCISCO J BEZANILLA; Fiscal Year: 2012
    ..different states of the channel, replacement of critical residues in the hydrophobic plug of the sensor, fluorescence polarization and anisotropy, molecular modeling and the crystal structures available...
  43. Novel Fluorophores for Molecular and Cellular Imaging
    Yihua Bruce Yu; Fiscal Year: 2013
    ..detection to eliminate background signal and study dynamics of molecular processes and interactions by fluorescence polarization and FRET...
  44. Novel Extended Red-Shifted Rhodamine Dyes
    NICHOLAS CAIRNS; Fiscal Year: 2004
    ..Assessment of the Performance of the New Dyes in a Model Fluorescence Polarization (FP) Study 5...
  45. HT screens for the disruption of the T cell receptor CD3 interface
    Luc Teyton; Fiscal Year: 2013
    ..We have created and optimized a fluorescence polarization assay amenable to high throughput screen using recombinant TCR molecules labeled with fluorophore on ..
  46. Chemical Tools for the Study of Dengue Virus Entry
    PRISCILLA LI NING YANG; Fiscal Year: 2011
    ..In Aim 1, we propose to use develop a high-throughput fluorescence polarization assay to identify ligands of the dengue virus envelope protein that inhibit dengue virus infection...
  47. HTS fluorescence polarization assay for inhibitors of Keap1-Nrf2 interaction
    Longqin Hu; Fiscal Year: 2012
    DESCRIPTION (provided by applicant): HTS fluorescence polarization assay for inhibitors of Keap1-Nrf2 interaction Keap1-Nrf2-antioxidant response element (ARE) system regulates cellular defense mechanisms that protect cells from ..
  48. MOLECULAR MECHANISMS OF BLOOD COAGULATION ACTIVATION
    JOSEPH SHORE; Fiscal Year: 1991
    ..These studies will use fluorescence polarization, fluorescence resonance energy transfer, enzyme kinetics and rapid reaction kinetic techniques...
  49. ER beta selective agonist coronary protection-menopause
    R Hermsmeyer; Fiscal Year: 2005
    ..PanVera ERbeta fluorescence polarization analysis adds genomic regulation dimensions to the plan...
  50. SUBUNIT INTERACTIONS IN DNA BINDING PROTEINS
    Catherine Royer; Fiscal Year: 1992
    ..Additionally, studies of the intrinsic tryptophan fluorescence polarization and lifetime, as a function of temperature, pressure, and ligation state will yield information as to ..
  51. OPTICAL STUDIES OF PROTEIN STRUCTURE AND FUNCTION
    LUBERT STRYER; Fiscal Year: 1991
    ..Rotation motions of domains will be delineated by nanosecond fluorescence polarization spectroscopy...
  52. Discovery of small-molecule inhibitors of the beta-catenin/BCL-9 interaction
    Shaomeng Wang; Fiscal Year: 2010
    ....
  53. Small-molecule inhibitors of the human RECQ1 and Bloom DNA helicases
    Opher Gileadi; Fiscal Year: 2009
    ..throughput, have been implemented for verification and classification: ATP binding, DNA binding (both by fluorescence polarization), and protein stabilization (thermal denaturation)...
  54. FUNCTION OF INITIATION FACTORS IN BACTERIAL TRANSLATION
    JOHN HERSHEY; Fiscal Year: 1992
    ..Third, the mechanism of action of initiation factors will be probed by in vitro mutagenesis and by fluorescence polarization techniques...
  55. IN VITRO STUDIES ON CALCIUM SENSITIVE ACTIN ASSEMBLY
    Joseph Bryan; Fiscal Year: 1991
    ..We now propose to use a variety of biochemical and biophysical methods, including fluorescence polarization and stopped flow techniques, to detail the interactions between Ca++, actin and gelsolin...
  56. STRUCTURAL BASIS OF KINESIN MOTOR MOVEMENT
    Hernando Sosa; Fiscal Year: 2006
    ..will be characterized at different points in the ATPase cycle using cryo- electron microscopy and fluorescence polarization microscopy on kinesin molecules fluorescently labeled on the motor domain...
  57. CANNABINOID EFFECTS ON MEMBRANES AND THEIR CONSTITUENTS
    Alan Bloom; Fiscal Year: 1990
    ..Studies of membrane fluidity using fluorescence polarization techniques will be continued...
  58. STRUCTURE-FUNCTION STUDIES OF HUMAN PLASMA FIBRONECTIN
    Robert Smith; Fiscal Year: 1990
    ..measurement of association constants between FN fragments and fluorphore-labeled gelatin fragment using fluorescence polarization in the fluid phase...
  59. Protein Dynamics in Ca2+-Regulated Thin Filaments
    Gerard Marriott; Fiscal Year: 2006
    ..measurements of fluorescence resonance energy transfer (FRET) efficiency, orientation factor KE2, fluorescence polarization (FP) on single, Ca2+-regulated filaments and by fitting these data into current models of the thin ..
  60. Fluorescent HTS Assay for Methyltransferases in Neurodegenerative Diseases
    Robert Lowery; Fiscal Year: 2006
    ..is based on detection of the invariable MT reaction product S-adenosyl homocysteine-using a competitive fluorescence polarization immunoassay...
  61. Preparation of New Long Wavelength Fluorescent Dyes
    NICHOLAS CAIRNS; Fiscal Year: 2005
    ..dyes by novel combinatorial chemistry techniques that are designed for use in high throughput screening, fluorescence polarization, biomedical imaging and other applicable techniques such as immunofluorescence microscopy and ..
  62. LIPID METABOLISM AND MEMBRANE STRUCTURE AND FUNCTION
    Michael Glaser; Fiscal Year: 1990
    ..of the membranes and reconstituted phospholipid vesicles containing membrane proteins will be studied by fluorescence polarization, energy transfer methods, and phase fluorometry...
  63. Protein synthesis on single ribosomes
    YALE GOLDMAN; Fiscal Year: 2004
    ..Two new methodological developments, single molecule fluorescence polarization and force-feedback infrared laser optical trap will be applied, providing information currently ..
  64. RHOMOCYCSTEINASE FOR HOMOCYSTEINE ASSAY
    Yuying Tan; Fiscal Year: 2002
    ..HPLC is highly specialized and low throughput, however. A fluorescence polarization immunoassay for tHCY has also been developed...
  65. Fluorescent HTS Assays for Methyltransferases in Neurodegenerative Diseases
    Robert Lowery; Fiscal Year: 2009
    ..To address this need, in Phase I BellBrook Labs developed a novel fluorescence polarization immunoassay for S-adenosylhomocysteine (SAH), the invariant product of all MT reactions...