fluorescence resonance energy transfer

Summary

Summary: A type of FLUORESCENCE SPECTROSCOPY using two FLUORESCENT DYES with overlapping emission and absorption spectra, which is used to indicate proximity of labeled molecules. This technique is useful for studying interactions of molecules and PROTEIN FOLDING.

Top Publications

  1. pmc Expanded dynamic range of fluorescent indicators for Ca(2+) by circularly permuted yellow fluorescent proteins
    Takeharu Nagai
    Laboratory for Cell Function and Dynamics, Advanced Technology Development Group, Brain Science Institute, RIKEN, 2 1 Hirosawa, Wako, Saitama 351 0198, Japan
    Proc Natl Acad Sci U S A 101:10554-9. 2004
  2. pmc A practical guide to single-molecule FRET
    Rahul Roy
    Department of Physics, University of Illinois at Urbana Champaign, 1110 West Green Street, Urbana, Illinois 61801, USA
    Nat Methods 5:507-16. 2008
  3. ncbi An improved cyan fluorescent protein variant useful for FRET
    Mark A Rizzo
    Nat Biotechnol 22:445-9. 2004
  4. pmc Development of an optimized backbone of FRET biosensors for kinases and GTPases
    Naoki Komatsu
    Laboratory of Bioimaging and Cell Signaling, Graduate School of Biostudies, Kyoto University, Kyoto 606 8501, Japan
    Mol Biol Cell 22:4647-56. 2011
  5. ncbi Rapid spontaneous accessibility of nucleosomal DNA
    Gu Li
    Department of Biochemistry, Molecular Biology, and Cell Biology, Northwestern University, Evanston, Illinois 60208 3500, USA
    Nat Struct Mol Biol 12:46-53. 2005
  6. pmc A genetically encoded fluorescent sensor of ERK activity
    Christopher D Harvey
    Janelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA
    Proc Natl Acad Sci U S A 105:19264-9. 2008
  7. pmc Visualization of ATP levels inside single living cells with fluorescence resonance energy transfer-based genetically encoded indicators
    Hiromi Imamura
    Precursory Research for Embryonic Science, Japan Science and Technology Agency, 5 Sanbancho, Chiyoda ku, Tokyo 102 0075, Japan
    Proc Natl Acad Sci U S A 106:15651-6. 2009
  8. ncbi Spatio-temporal Rho GTPase signaling - where are we now?
    Olivier Pertz
    Institute for Biochemistry and Genetics, Department of Biomedicine, University of Basel, Mattenstrasse 28, 4058 Basel, Switzerland
    J Cell Sci 123:1841-50. 2010
  9. pmc Analysis of single-molecule FRET trajectories using hidden Markov modeling
    Sean A McKinney
    Department of Physics and Howard Hughes Medical Institute, University of Illinois at Urbana Champaign, Urbana, Illinois, USA
    Biophys J 91:1941-51. 2006
  10. pmc Probing single-stranded DNA conformational flexibility using fluorescence spectroscopy
    M C Murphy
    Physics Department, University of Illinois at Urbana Champaign, Urbana, Illinois 61801, USA
    Biophys J 86:2530-7. 2004

Detail Information

Publications336 found, 100 shown here

  1. pmc Expanded dynamic range of fluorescent indicators for Ca(2+) by circularly permuted yellow fluorescent proteins
    Takeharu Nagai
    Laboratory for Cell Function and Dynamics, Advanced Technology Development Group, Brain Science Institute, RIKEN, 2 1 Hirosawa, Wako, Saitama 351 0198, Japan
    Proc Natl Acad Sci U S A 101:10554-9. 2004
    b>Fluorescence resonance energy transfer (FRET) technology has been used to develop genetically encoded fluorescent indicators for various cellular functions...
  2. pmc A practical guide to single-molecule FRET
    Rahul Roy
    Department of Physics, University of Illinois at Urbana Champaign, 1110 West Green Street, Urbana, Illinois 61801, USA
    Nat Methods 5:507-16. 2008
    Single-molecule fluorescence resonance energy transfer (smFRET) is one of the most general and adaptable single-molecule techniques...
  3. ncbi An improved cyan fluorescent protein variant useful for FRET
    Mark A Rizzo
    Nat Biotechnol 22:445-9. 2004
    ..Cerulean is 2.5-fold brighter than ECFP and replacement of ECFP with Cerulean substantially improves the signal-to-noise ratio of a FRET-based sensor for glucokinase activation...
  4. pmc Development of an optimized backbone of FRET biosensors for kinases and GTPases
    Naoki Komatsu
    Laboratory of Bioimaging and Cell Signaling, Graduate School of Biostudies, Kyoto University, Kyoto 606 8501, Japan
    Mol Biol Cell 22:4647-56. 2011
    ..Furthermore, this backbone enabled us to develop novel FRET biosensors for several kinases of RSK, S6K, Akt, and PKC and to perform quantitative evaluation of kinase inhibitors in living cells...
  5. ncbi Rapid spontaneous accessibility of nucleosomal DNA
    Gu Li
    Department of Biochemistry, Molecular Biology, and Cell Biology, Northwestern University, Evanston, Illinois 60208 3500, USA
    Nat Struct Mol Biol 12:46-53. 2005
    ....
  6. pmc A genetically encoded fluorescent sensor of ERK activity
    Christopher D Harvey
    Janelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA
    Proc Natl Acad Sci U S A 105:19264-9. 2008
    ..EKAR therefore permits the measurement of spatiotemporal ERK signaling dynamics in living cells, including in neuronal compartments in intact tissues...
  7. pmc Visualization of ATP levels inside single living cells with fluorescence resonance energy transfer-based genetically encoded indicators
    Hiromi Imamura
    Precursory Research for Embryonic Science, Japan Science and Technology Agency, 5 Sanbancho, Chiyoda ku, Tokyo 102 0075, Japan
    Proc Natl Acad Sci U S A 106:15651-6. 2009
    ..To visualize ATP levels, we generated a series of fluorescence resonance energy transfer (FRET)-based indicators for ATP that were composed of the epsilon subunit of the bacterial F(o)F(1)..
  8. ncbi Spatio-temporal Rho GTPase signaling - where are we now?
    Olivier Pertz
    Institute for Biochemistry and Genetics, Department of Biomedicine, University of Basel, Mattenstrasse 28, 4058 Basel, Switzerland
    J Cell Sci 123:1841-50. 2010
    ..The next challenge is to decipher the molecular circuitry underlying this complex spatio-temporal modularity to produce integrated models of Rho GTPase signaling...
  9. pmc Analysis of single-molecule FRET trajectories using hidden Markov modeling
    Sean A McKinney
    Department of Physics and Howard Hughes Medical Institute, University of Illinois at Urbana Champaign, Urbana, Illinois, USA
    Biophys J 91:1941-51. 2006
    The analysis of single-molecule fluorescence resonance energy transfer (FRET) trajectories has become one of significant biophysical interest...
  10. pmc Probing single-stranded DNA conformational flexibility using fluorescence spectroscopy
    M C Murphy
    Physics Department, University of Illinois at Urbana Champaign, Urbana, Illinois 61801, USA
    Biophys J 86:2530-7. 2004
    ..The persistence length thus estimated ranges from 1.5 nm in 2 M NaCl to 3 nm in 25 mM NaCl...
  11. ncbi Coupling of ribosomal L1 stalk and tRNA dynamics during translation elongation
    Jingyi Fei
    Department of Chemistry, Columbia University, New York, NY 10027, USA
    Mol Cell 30:348-59. 2008
    By using single-molecule fluorescence resonance energy transfer (smFRET), we observe the real-time dynamic coupling between the ribosome, labeled at the L1 stalk, and transfer RNA (tRNA)...
  12. pmc Conversion of red fluorescent protein into a bright blue probe
    Oksana M Subach
    Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA
    Chem Biol 15:1116-24. 2008
    ....
  13. pmc Homomeric and heteromeric assembly of KCNQ (Kv7) K+ channels assayed by total internal reflection fluorescence/fluorescence resonance energy transfer and patch clamp analysis
    Manjot Bal
    Department of Physiology, University of Texas Health Science Center, San Antonio, Texas 78229, USA
    J Biol Chem 283:30668-76. 2008
    ..explore the heteromeric assembly of KCNQ channels in individual living cells, we performed fluorescence resonance energy transfer (FRET) between cyan fluorescent protein- and yellow fluorescent protein-tagged KCNQ subunits ..
  14. pmc Extreme conformational diversity in human telomeric DNA
    J Y Lee
    Department of Physics, Seoul National University, Seoul 151 742, Korea
    Proc Natl Acad Sci U S A 102:18938-43. 2005
    ..The extremely diverse conformations of the human telomeric DNA may have mechanistic implications for the proteins and drugs that recognize G-rich sequences...
  15. pmc SSB protein diffusion on single-stranded DNA stimulates RecA filament formation
    Rahul Roy
    Center for Biophysics and Computational Biology, University of Illinois, Urbana Champaign, Illinois 61801, USA
    Nature 461:1092-7. 2009
    ..Here we show, using single-molecule two- and three-colour fluorescence resonance energy transfer, that tetrameric SSB can spontaneously migrate along ssDNA...
  16. ncbi Advances in single-molecule fluorescence methods for molecular biology
    Chirlmin Joo
    Department of Physics, University of Illinois at Urbana Champaign, Urbana, IL 61801, USA
    Annu Rev Biochem 77:51-76. 2008
    ..In this exciting era of transitioning from in vitro to in vivo and in situ conditions, it is anticipated that SM fluorescence methodology will become a common tool of molecular biology...
  17. ncbi Measuring calcium signaling using genetically targetable fluorescent indicators
    Amy E Palmer
    Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado 80309 0215, USA
    Nat Protoc 1:1057-65. 2006
    ..An experiment and calibration will probably take a few hours. Finally, the offline data workup can take approximately 1 d depending on the extent of analysis...
  18. ncbi The fluorescent toolbox for assessing protein location and function
    Ben N G Giepmans
    National Center for Microscopy and Imaging Research, Center for Research in Biological Systems, Department of Neurosciences, University of California, San Diego, La Jolla, CA 92093, USA
    Science 312:217-24. 2006
    ..Small organic fluorescent dyes, nanocrystals ("quantum dots"), autofluorescent proteins, small genetic encoded tags that can be complexed with fluorochromes, and combinations of these probes are highlighted...
  19. pmc Spontaneous intersubunit rotation in single ribosomes
    Peter V Cornish
    Department of Physics, University of Illinois, 1110 West Green Street, Urbana, IL 61801, USA
    Mol Cell 30:578-88. 2008
    ..Our results provide direct evidence that the intersubunit rotation that underlies ribosomal translocation is thermally driven...
  20. ncbi Creating new fluorescent probes for cell biology
    Jin Zhang
    Department of Pharmacology, Massachusetts Institute of Technology, 77 Massachusetts Avenue, 18 496, Cambridge, Massachusetts 02139, USA
    Nat Rev Mol Cell Biol 3:906-18. 2002
    ....
  21. ncbi In vivo fluorescence resonance energy transfer imaging reveals differential activation of Rho-family GTPases in glioblastoma cell invasion
    Eishu Hirata
    Department of Bioimaging and Cell Signaling, Graduate School of Biostudies, Kyoto University, Kyoto, 606 8501, Japan
    J Cell Sci 125:858-68. 2012
    ....
  22. pmc Split-protein systems: beyond binary protein-protein interactions
    Sujan S Shekhawat
    Department of Chemistry and Biochemistry, University of Arizona, 1306 East University Blvd, Tucson, AZ 85721, USA
    Curr Opin Chem Biol 15:789-97. 2011
    ..Finally, we discuss autoinhibition strategies leading to turn-on sensors as well as future directions in split-protein methodology including possible therapeutic approaches...
  23. ncbi A toolkit and benchmark study for FRET-restrained high-precision structural modeling
    Stanislav Kalinin
    Lehrstuhl fur Molekulare Physikalische Chemie, Heinrich Heine Universität HHU, Dusseldorf, Germany
    Nat Methods 9:1218-25. 2012
    ..We used this hybrid approach to determine the formerly unknown configuration of the flexible single-strand template overhang...
  24. pmc Defining the limits of single-molecule FRET resolution in TIRF microscopy
    Seamus J Holden
    Biological Physics Research Group, Department of Physics, University of Oxford, Oxford, United Kingdom
    Biophys J 99:3102-11. 2010
    ..the mechanisms of which are often characterized by distance changes on the scale of one DNA basepair...
  25. ncbi Quantum dot-aptamer conjugates for synchronous cancer imaging, therapy, and sensing of drug delivery based on bi-fluorescence resonance energy transfer
    Vaishali Bagalkot
    Laboratory of Nanomedicine and Biomaterials, Department of Anesthesiology, Brigham and Women s Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA
    Nano Lett 7:3065-70. 2007
    ..A donor-acceptor model fluorescence resonance energy transfer (FRET) between QD and Dox and a donor-quencher model FRET between Dox and aptamer result when Dox ..
  26. ncbi Fluorescent protein FRET pairs for ratiometric imaging of dual biosensors
    Hui Wang Ai
    University of Alberta, Department of Chemistry, 11227 Saskatchewan Drive, Edmonton, Alberta T6G 2G2, Canada
    Nat Methods 5:401-3. 2008
    b>Fluorescence resonance energy transfer (FRET) with fluorescent proteins is a powerful method for detection of protein-protein interactions, enzyme activities and small molecules in the intracellular milieu...
  27. pmc Conformational trapping of mismatch recognition complex MSH2/MSH3 on repair-resistant DNA loops
    Walter H Lang
    Lawrence Berkeley National Laboratory, Life Sciences Division, 1 Cyclotron Road, Berkeley, CA 94720, USA
    Proc Natl Acad Sci U S A 108:E837-44. 2011
    ..We envision that junction dynamics is an active participant and a conformational regulator of repair signaling, and governs whether a loop is removed by MSH2/MSH3 or escapes to become a precursor for mutation...
  28. pmc Initial transcription by RNA polymerase proceeds through a DNA-scrunching mechanism
    Achillefs N Kapanidis
    Department of Chemistry and Biochemistry and Department of Physiology, University of California, Los Angeles, CA 90095, USA
    Science 314:1144-7. 2006
    Using fluorescence resonance energy transfer to monitor distances within single molecules of abortively initiating transcription initiation complexes, we show that initial transcription proceeds through a "scrunching" mechanism, in which ..
  29. pmc Nucleosome accessibility governed by the dimer/tetramer interface
    Vera Böhm
    Abteilung Biophysik der Makromolekule, Deutsches Krebsforschungszentrum, Heidelberg, Germany
    Nucleic Acids Res 39:3093-102. 2011
    ..2-3% under physiological conditions. This finding could have significant in vivo implications for factor-mediated histone removal and exchange, as well as for regulating DNA accessibility to the transcription and replication machinery...
  30. pmc Cerulean, Venus, and VenusY67C FRET reference standards
    Srinagesh V Koushik
    Laboratory of Molecular Physiology, National Institute on Alcohol Abuse and Alcoholism, Bethesda, MD, USA
    Biophys J 91:L99-L101. 2006
    ..Since the results from each method were in good agreement, the FRET efficiency value of each construct could be determined with high accuracy and precision, thereby justifying their use as standards...
  31. pmc Interplay of alpha-synuclein binding and conformational switching probed by single-molecule fluorescence
    Allan Chris M Ferreon
    Department of Molecular Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA
    Proc Natl Acad Sci U S A 106:5645-50. 2009
    ..Using single-molecule fluorescence resonance energy transfer and correlation methods, we directly probed protein membrane association, structural distributions,..
  32. pmc Imaging of the Yellow Cameleon 3.6 indicator reveals that elevations in cytosolic Ca2+ follow oscillating increases in growth in root hairs of Arabidopsis
    Gabriele B Monshausen
    Department of Botany, University of Wisconsin, Madison, Wisconsin 53706, USA
    Plant Physiol 147:1690-8. 2008
    ..6 fluorescence resonance energy transfer-based Ca(2+) sensor...
  33. ncbi Cyan and yellow super fluorescent proteins with improved brightness, protein folding, and FRET Förster radius
    Gert Jan Kremers
    Section Molecular Cytology and Centre for Advanced Microscopy, Swammerdam Institute for Life Sciences, University of Amsterdam, Kruislaan 316, 1098 SM, Amsterdam, The Netherlands
    Biochemistry 45:6570-80. 2006
    ..proteins are widely used for dual color imaging and protein-protein interaction studies based on fluorescence resonance energy transfer. Use of these fluorescent proteins can be limited by their thermosensitivity, dim fluorescence, ..
  34. 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...
  35. ncbi Illuminating insights into protein-protein interactions using bioluminescence resonance energy transfer (BRET)
    Kevin D G Pfleger
    7TM Laboratory Laboratory for Molecular Endocrinology, Western Australian Institute for Medical Research, UWA Centre for Medical Research, University of Western Australia, Nedlands, Perth, Western Australia 6009, Australia
    Nat Methods 3:165-74. 2006
    ..Therefore, this review aims to demystify and clarify the important aspects of the BRET methodology that should be considered when using this technique...
  36. ncbi Formation of tight junction: determinants of homophilic interaction between classic claudins
    Jörg Piontek
    Leibniz Institut fur Molekulare Pharmakologie, Robert Rossle Str 10, 13125 Berlin, Germany
    FASEB J 22:146-58. 2008
    ..Since nearly all these residues are conserved in most claudins, our findings are of general relevance for all classical claudins. On the basis of the data we have established a novel molecular concept for tight junction formation...
  37. pmc Efficiently folding and circularly permuted variants of the Sapphire mutant of GFP
    Otilia Zapata-Hommer
    Max Planck Institut für Neurobiologie Am Klopferspitz 18a 82152 Martinsried, Germany
    BMC Biotechnol 3:5. 2003
    ..gene has yielded a number of improved variants such as EGFP or colour variants suitable for fluorescence resonance energy transfer (FRET)...
  38. ncbi Genetically encoded FRET sensors to monitor intracellular Zn2+ homeostasis
    Jan L Vinkenborg
    Laboratory of Chemical Biology, Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands
    Nat Methods 6:737-40. 2009
    We developed genetically encoded fluorescence resonance energy transfer (FRET)-based sensors that display a large ratiometric change upon Zn(2+) binding, have affinities that span the pico- to nanomolar range and can readily be targeted ..
  39. pmc Determination of hierarchical relationship of Src and Rac at subcellular locations with FRET biosensors
    Mingxing Ouyang
    Department of Bioengineering, Beckman Institute for Advanced Science and Technology, University of Illinois, Urbana Champaign, IL 61801, USA
    Proc Natl Acad Sci U S A 105:14353-8. 2008
    ..In summary, ECFP/YPet may serve as a general FRET pair for the development of highly sensitive biosensors to allow the determination of molecular hierarchies at subcellular locations in live cells...
  40. pmc Measuring steady-state and dynamic endoplasmic reticulum and Golgi Zn2+ with genetically encoded sensors
    Yan Qin
    Department of Chemistry and Biochemistry, University of Colorado, Boulder, CO 80309, USA
    Proc Natl Acad Sci U S A 108:7351-6. 2011
    ..This study provides direct evidence that Ca(2+) signaling can influence Zn(2+) homeostasis and vice versa, that Zn(2+) dynamics may modulate Ca(2+) signaling...
  41. ncbi Imaging protein activity in live embryos using fluorescence resonance energy transfer biosensors
    Elena Kardash
    Institute for Cell Biology, Center for Molecular Biology of Inflammation, University of Munster, Germany
    Nat Protoc 6:1835-46. 2011
    b>Fluorescence resonance energy transfer (FRET)-based molecular biosensors serve as important tools for studying protein activity in live cells and have been widely used for this purpose over the past decade...
  42. ncbi Visualizing the mechanical activation of Src
    Yingxiao Wang
    Department of Bioengineering and The Whitaker Institute of Biomedical Engineering, La Jolla, California 92093, USA
    Nature 434:1040-5. 2005
    ..We find that the transmission of mechanically induced Src activation is a dynamic process that directs signals via the cytoskeleton to spatial destinations...
  43. pmc mRNA translocation occurs during the second step of ribosomal intersubunit rotation
    Dmitri N Ermolenko
    Center for Molecular Biology of RNA, University of California, Santa Cruz, Santa Cruz, California, USA
    Nat Struct Mol Biol 18:457-62. 2011
    ..combination of two fluorescence-based approaches to study the timing of these events, intersubunit fluorescence resonance energy transfer measurements to observe relative rotational movement of the subunits, and a fluorescence quenching ..
  44. pmc Single-molecule Förster resonance energy transfer reveals an innate fidelity checkpoint in DNA polymerase I
    Svitlana Y Berezhna
    Department of Molecular Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA
    J Am Chem Soc 134:11261-8. 2012
    ..These results reveal the existence of an early innate fidelity checkpoint, rejecting incorrect nucleotide substrates before the enzyme encloses the nascent base pair...
  45. pmc Three-color spectral FRET microscopy localizes three interacting proteins in living cells
    Yuansheng Sun
    W M Keck Center for Cellular Imaging, Departments of Biology and Biomedical Engineering, University of Virginia, Charlottesville, Virginia, USA
    Biophys J 99:1274-83. 2010
    ..We show how the 3sFRET microscopy method represents a promising live-cell imaging technique to monitor the interactions between three labeled cellular components...
  46. ncbi Conformational changes of DEAD-box helicases monitored by single molecule fluorescence resonance energy transfer
    Alexandra Z Andreou
    Institute for Physical Chemistry, University of Muenster, Muenster, Germany
    Methods Enzymol 511:75-109. 2012
    ..Here, we describe single molecule fluorescence resonance energy transfer (smFRET) approaches to directly monitor conformational changes associated with opening and closing ..
  47. pmc Natural Leishmania infection of Lutzomyia auraensis in Madre de Dios, Peru, detected by a fluorescence resonance energy transfer-based real-time polymerase chain reaction
    Hugo O Valdivia
    United States Naval Medical Research Unit No 6, Lima, Peru
    Am J Trop Med Hyg 87:511-7. 2012
    ..auraensis was estimated to be 0.6% (95% confidence interval = 0.20-1.42%). Further studies are needed to assess the importance of Lu. auraensis in the transmission of New World tegumentary leishmaniasis in hyperendemic areas of Peru...
  48. ncbi Amine-functionalized lanthanide-doped zirconia nanoparticles: optical spectroscopy, time-resolved fluorescence resonance energy transfer biodetection, and targeted imaging
    Yongsheng Liu
    Key Laboratory of Optoelectronic Materials Chemistry and Physics, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, Fujian 350002, China
    J Am Chem Soc 134:15083-90. 2012
    ..the long-lived luminescence of Ln(3+), we demonstrate its application as a sensitive time-resolved fluorescence resonance energy transfer (FRET) bioprobe to detect avidin with a record-low detection limit of 3.0 nM...
  49. pmc Orientation of cyanine fluorophores terminally attached to DNA via long, flexible tethers
    Jonathan Ouellet
    Cancer Research UK Nucleic Acid Structure Research Group, The University of Dundee, Dundee, United Kingdom
    Biophys J 101:1148-54. 2011
    ..This is consistent with the transition moments becoming approximately aligned with the long axis of the terminal basepair for the long-linker species...
  50. pmc STIM1-Orai1 interactions and Orai1 conformational changes revealed by live-cell FRET microscopy
    Laura Navarro-Borelly
    Department of Molecular Pharmacology and Biological Chemistry, Northwestern University, Feinberg School of Medicine, 303 E Chicago Avenue, Ward 8 296, Chicago, IL 60611, USA
    J Physiol 586:5383-401. 2008
    ..we show that redistribution of STIM1 and Orai1 occurs in parallel with a pronounced increase in fluorescence resonance energy transfer (FRET) between STIM1 and Orai1, supporting the idea that activation of CRAC channels occurs ..
  51. pmc Time-resolved fluorescence resonance energy transfer (TR-FRET) to analyze the disruption of EGFR/HER2 dimers: a new method to evaluate the efficiency of targeted therapy using monoclonal antibodies
    Nadège Gaborit
    IRCM, Institut de Recherche en Cancérologie de Montpellier, INSERM, U896, Université Montpellier1, CRLC Val d Aurelle Paul Lamarque, Montpellier, France
    J Biol Chem 286:11337-45. 2011
    ..Here, we describe a time-resolved fluorescence resonance energy transfer (TR-FRET) method to quantify EGFR/HER2 heterodimers on cell surface to shed some light on the ..
  52. pmc Following movement of the L1 stalk between three functional states in single ribosomes
    Peter V Cornish
    Department of Physics, University of Illinois, 1110 West Green Street, Urbana, IL 61801, USA
    Proc Natl Acad Sci U S A 106:2571-6. 2009
    ....
  53. pmc Dynamics of nucleosome remodelling by individual ACF complexes
    Timothy R Blosser
    Howard Hughes Medical Institute, Massachusetts 02138, USA
    Nature 462:1022-7. 2009
    ..At equilibrium, a continuously bound ACF complex can move the nucleosome back-and-forth many times before dissociation, indicating that ACF is a highly processive and bidirectional nucleosome translocase...
  54. pmc GLUT1 and GLUT9 as major contributors to glucose influx in HepG2 cells identified by a high sensitivity intramolecular FRET glucose sensor
    Hitomi Takanaga
    Carnegie Institution for Science, 260 Panama Street, Stanford CA 94305, USA
    Biochim Biophys Acta 1778:1091-9. 2008
    ..The improved sensitivity of the novel glucose nanosensor increases the reliability of in vivo glucose flux analyses, and provides a new means for the screening of siRNA collections as well as drugs using high-content screens...
  55. ncbi Improving membrane voltage measurements using FRET with new fluorescent proteins
    Hidekazu Tsutsui
    Laboratory for Cell Function Dynamics, Brain Science Institute, RIKEN, 2 1 Hirosawa, Saitama 351 0198, Japan
    Nat Methods 5:683-5. 2008
    We used two new coral fluorescent proteins as fluorescence resonance energy transfer (FRET) donor and acceptor to develop a voltage sensor, named Mermaid, that displays approximately 40% changes in emission ratio per 100 mV, allowing for ..
  56. pmc Highly sensitive and quantitative FRET-FLIM imaging in single dendritic spines using improved non-radiative YFP
    Hideji Murakoshi
    Department of Neurobiology, Duke University Medical Center, Durham, NC 27710, USA
    Brain Cell Biol 36:31-42. 2008
    ..fluorescence lifetime imaging microscopy (TPFLIM) enables the quantitative measurements of fluorescence resonance energy transfer (FRET) in small subcellular compartments in light scattering tissue...
  57. ncbi In vivo imaging of the dynamics of glucose uptake in the cytosol of COS-7 cells by fluorescent nanosensors
    Marcus Fehr
    Zentrum für Molekular biologie der Pflanzen Tübingen, Plant Physiology, Auf der Morgenstelle 1, D 72076 Tubingen, Germany
    J Biol Chem 278:19127-33. 2003
    ..Upon binding of substrate the FLIPglu-170n sensor showed a concentration-dependent decrease in fluorescence resonance energy transfer between the attached chromophores with a binding affinity for glucose of 170 nm...
  58. ncbi Dynamics of the Ras/ERK MAPK cascade as monitored by fluorescent probes
    Aki Fujioka
    Department of Signal Transduction, Research Institute for Microbial Diseases, Osaka University, Yamadaoka, Suita Shi, Osaka 565 0871, Japan
    J Biol Chem 281:8917-26. 2006
    ..the activation and nuclear translocation of ERK, we developed probes based on the principle of fluorescence resonance energy transfer. Third, the dissociation constants of Ras.Raf, Raf.MEK, and MEK...
  59. ncbi Graphical representation and multicomponent analysis of single-frequency fluorescence lifetime imaging microscopy data
    A H A Clayton
    Ludwig Institute for Cancer Research, PO Box 2008, Royal Melbourne Hospital, Parkville, Victoria 3050, Australia
    J Microsc 213:1-5. 2004
    ..The graphical approach was applied to cellular data obtained from a lifetime-based phosphorylation assay for the epidermal growth factor receptor and yielded results similar to those obtained by a global analysis algorithm...
  60. ncbi Nanostructured probes for RNA detection in living cells
    Philip Santangelo
    Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA
    Ann Biomed Eng 34:39-50. 2006
    ..In particular, we describe methods that use dual FRET (fluorescence resonance energy transfer) or single molecular beacons in combination with peptide-based or membrane-permeabilization-based ..
  61. pmc Fluorescence-aided molecule sorting: analysis of structure and interactions by alternating-laser excitation of single molecules
    Achillefs N Kapanidis
    Department of Chemistry and Biochemistry, University of California, 607 Charles E Young Drive East, Los Angeles, CA 90095, USA
    Proc Natl Acad Sci U S A 101:8936-41. 2004
    ....
  62. ncbi Visualization of PKA activity in plasma membrane microdomains
    CHARLENE DEPRY
    Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA
    Mol Biosyst 7:52-8. 2011
    ....
  63. pmc An improved cerulean fluorescent protein with enhanced brightness and reduced reversible photoswitching
    Michele L Markwardt
    Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland, United States of America
    PLoS ONE 6:e17896. 2011
    ..Thus, mCerulean3 is a bright, photostable cyan fluorescent protein which possesses several characteristics that are highly desirable for FRET experiments...
  64. ncbi Spatio-temporal regulation of Rac1 and Cdc42 activity during nerve growth factor-induced neurite outgrowth in PC12 cells
    Kazuhiro Aoki
    Department of Tumor Virology, Research Institute for Microbial Diseases, Osaka University, Yamadaoka, Suita Shi, Osaka 565 0871, Japan
    J Biol Chem 279:713-9. 2004
    ..NGF)-induced neurite outgrowth in living PC12 cells by using probes based on the principle of fluorescence resonance energy transfer (FRET)...
  65. pmc Learning rates and states from biophysical time series: a Bayesian approach to model selection and single-molecule FRET data
    Jonathan E Bronson
    Department of Chemistry, Columbia University, New York, New York, USA
    Biophys J 97:3196-205. 2009
    ..Source code used in this analysis, including a graphical user interface, is available open source via http://vbFRET.sourceforge.net...
  66. pmc Conformational transitions in DNA polymerase I revealed by single-molecule FRET
    Yusdi Santoso
    Biological Physics Research Group, Department of Physics, Clarendon Laboratory, University of Oxford, Oxford OX1 3PU, United Kingdom
    Proc Natl Acad Sci U S A 107:715-20. 2010
    ..fragment) labeled with both donor and acceptor fluorophores, we have employed single-molecule fluorescence resonance energy transfer to study the polymerase conformational transitions that precede nucleotide addition...
  67. pmc Structure and dynamics of a processive Brownian motor: the translating ribosome
    Joachim Frank
    Howard Hughes Medical Institute, Department of Biochemistry and Molecular Biophysics, Columbia University, New York City, New York 10032, USA
    Annu Rev Biochem 79:381-412. 2010
    ..microscopic and X-ray crystallographic structural studies, on the one hand, and single-molecule fluorescence resonance energy transfer (smFRET) dynamic studies, on the other, is emerging as a powerful means for investigating the ..
  68. pmc Fluorescence resonance energy transfer-based stoichiometry in living cells
    Adam Hoppe
    Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI 48109, USA
    Biophys J 83:3652-64. 2002
    Imaging of fluorescence resonance energy transfer (FRET) between fluorescently labeled molecules can measure the timing and location of intermolecular interactions inside living cells...
  69. pmc Investigation of the dimerization of proteins from the epidermal growth factor receptor family by single wavelength fluorescence cross-correlation spectroscopy
    Ping Liu
    Department of Chemistry, National University of Singapore, Singapore
    Biophys J 93:684-98. 2007
    ..These receptors are major targets of anti-cancer drug development, and the receptors' homo- and heterodimeric structures are relevant for such developments...
  70. ncbi Fluorescence lifetime and polarization-resolved imaging in cell biology
    James A Levitt
    Department of Physics, King s College London, London, UK
    Curr Opin Biotechnol 20:28-36. 2009
    ..The particular merits of polarization-resolved fluorescence measurements are highlighted, including the unique ability to elucidate the occurrence of homo-FRET...
  71. pmc FRET or no FRET: a quantitative comparison
    Claude Berney
    BioMicroMetrics Group, Laboratory for Biomechanics, Swiss Federal Institute of Technology, CH 8952 Schlieren, Switzerland
    Biophys J 84:3992-4010. 2003
    b>Fluorescence resonance energy transfer (FRET) is a technique used to measure the interaction between two molecules labeled with two different fluorophores (the donor and the acceptor) by the transfer of energy from the excited donor to ..
  72. pmc Identification of pregnane X receptor ligands using time-resolved fluorescence resonance energy transfer and quantitative high-throughput screening
    Sunita J Shukla
    NIH Chemical Genomics Center, National Institutes of Health, Bethesda, Maryland, USA
    Assay Drug Dev Technol 7:143-69. 2009
    ..To facilitate the identification of PXR ligands, a time-resolved fluorescence resonance energy transfer (TR-FRET) assay was miniaturized to a 1,536-well microtiter plate format to employ quantitative ..
  73. ncbi TFIIA changes the conformation of the DNA in TBP/TATA complexes and increases their kinetic stability
    Aaron R Hieb
    Department of Chemistry and Biochemistry, University of Colorado at Boulder, 215 UCB, Boulder, CO 80309 0215, USA
    J Mol Biol 372:619-32. 2007
    ..We have used quantitative solution fluorescence resonance energy transfer (FRET) and gel-based FRET (gelFRET) to determine the effect of TFIIA on the conformation of the ..
  74. pmc Dynamic structural rearrangements between DNA binding modes of E. coli SSB protein
    Rahul Roy
    Center for Biophysics and Computational Biology, University of Illinois, Urbana Champaign, IL 61801, USA
    J Mol Biol 369:1244-57. 2007
    ....
  75. pmc Structure of the three-way helical junction of the hepatitis C virus IRES element
    Jonathan Ouellet
    Cancer Research UK Nucleic Acid Structure Research Group, MSI WTB complex, The University of Dundee, Dundee DD1 5EH, United Kingdom
    RNA 16:1597-609. 2010
    ..Comparative gel electrophoresis and ensemble fluorescence resonance energy transfer (FRET) studies show that the junction is induced to fold by the presence of Mg(2+) ions in low ..
  76. ncbi Fluorescence resonance energy transfer (FRET)-based biosensors: visualizing cellular dynamics and bioenergetics
    Sohila Zadran
    David Geffen School of Medicine, University of California, Los Angeles, CA, USA
    Appl Microbiol Biotechnol 96:895-902. 2012
    ..Here, we will review current advances and limitations of FRET-based biosensor technology and discuss future FRET applications...
  77. ncbi Spatiotemporal regulation of small GTPases as revealed by probes based on the principle of Förster Resonance Energy Transfer (FRET): Implications for signaling and pharmacology
    Etsuko Kiyokawa
    Department of Pathology and Biology of Diseases, Kyoto University, Japan
    Annu Rev Pharmacol Toxicol 51:337-58. 2011
    ..In the process, we attempt to delineate the merits, current limitations, and future applications of this technique to pharmacological studies...
  78. ncbi G protein-coupled receptor heteromer identification technology: identification and profiling of GPCR heteromers
    Sanam Mustafa
    Laboratory for Molecular Endocrinology GPCRs, Western Australian Institute for Medical Research WAIMR and Centre for Medical Research, University of Western Australia, Nedlands, Western Australia, Australia
    J Lab Autom 16:285-91. 2011
    ..In this innovation brief, a novel technology developed to identify and profile GPCR heteromers and their ligands will be reviewed...
  79. ncbi Directional migration of neural crest cells in vivo is regulated by Syndecan-4/Rac1 and non-canonical Wnt signaling/RhoA
    Helen K Matthews
    Department of Anatomy and Development Biology, University College London, Gower Street, London WC1E 6BT, UK
    Development 135:1771-80. 2008
    ..In addition, we show that RhoA inhibits Rac in NC cells. We present a model in which Syn4 and PCP control directional NC migration by, at least in part, regulating membrane protrusions through the regulation of small GTPase activities...
  80. pmc G protein-coupled receptors sense fluid shear stress in endothelial cells
    Mirianas Chachisvilis
    La Jolla Bioengineering Institute, 505 Coast Boulevard South, La Jolla, CA 92037, USA
    Proc Natl Acad Sci U S A 103:15463-8. 2006
    ..Furthermore, because GPCRs are a major target of drug development, a detailed characterization of mechanochemical signaling via the GPCR pathway will be relevant for the development of new antiatherosclerosis drugs...
  81. pmc Localization of the dantrolene-binding sequence near the FK506-binding protein-binding site in the three-dimensional structure of the ryanodine receptor
    Ruiwu Wang
    Libin Cardiovascular Institute of Alberta, Department of Physiology and Pharmacology, University of Calgary, Calgary, Alberta T2N 4N1, Canada
    J Biol Chem 286:12202-12. 2011
    ..6-binding site but distant to the central region around residue Ser-2367. An allosteric mechanism by which dantrolene stabilizes interdomain interactions between the NH2-terminal and central regions is proposed...
  82. ncbi A nano-positioning system for macromolecular structural analysis
    Adam Muschielok
    Department of Chemistry and Biochemistry and Center for Integrated Protein Science München, Ludwig Maximilians Universitat Munchen, Butenandtstrasse 11, 81377 Munchen, Germany
    Nat Methods 5:965-71. 2008
    ..With this approach, single-pair fluorescence resonance energy transfer measurements can now be used as a quantitative tool for investigating the position and dynamics of ..
  83. ncbi Molecular basis of RNA-mediated gene regulation on the adenine riboswitch by single-molecule approaches
    Jean François Lemay
    Departement de Biologie, Universite de Sherbrooke, Quebec, Canada J1K 2R1
    Methods Mol Biol 540:65-76. 2009
    ..b>Fluorescence resonance energy transfer (FRET) and single-molecule FRET studies demonstrate the ability of the loops to interact in the ..
  84. pmc Pulsed interleaved excitation
    Barbara K Müller
    Physical Chemistry, Department of Chemistry and Biochemistry, and Center for NanoScience, Ludwig Maximilians Universitat Munchen, Butenandstrasse 11 Haus E, D 81377 Munich, Germany
    Biophys J 89:3508-22. 2005
    ..fluorescence imaging, fluorescence cross-correlation spectroscopy (FCCS), and single-pair fluorescence resonance energy transfer (spFRET) measurements...
  85. pmc Oligomerization is crucial for the stability and function of heme oxygenase-1 in the endoplasmic reticulum
    Hsuan Wen Hwang
    Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan
    J Biol Chem 284:22672-9. 2009
    ..Using chemical cross-linking, co-immunoprecipitation, and fluorescence resonance energy transfer (FRET) experiments, here we showed that HO-1 forms dimers/oligomers in the ER...
  86. ncbi Dynamic regulation of CXCR1 and CXCR2 homo- and heterodimers
    Laura Martínez Muñoz
    Department of Immunology and Oncology, Centro Nacional de Biotecnología Consejo Superior de Investigaciones Científicas, Cantoblanco, Madrid, Spain
    J Immunol 183:7337-46. 2009
    ..neutrophils and in cell lines that coexpress the chemokine receptors CXCR1 and CXCR2, we used fluorescence resonance energy transfer techniques to show that these two receptors form homo- and heterodimers...
  87. pmc Genetically encoded probe for fluorescence lifetime imaging of CaMKII activity
    Showming Kwok
    RIKEN MIT Neuroscience Research Center, The Picower Institute for Learning and Memory, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, 77 Massachusetts Avenue 46 4243A, Cambridge, MA 02139, USA
    Biochem Biophys Res Commun 369:519-25. 2008
    ..These FLIM versions of Camui could be useful for elucidating the function of CaMKII both in vitro and in vivo...
  88. pmc Allosteric collaboration between elongation factor G and the ribosomal L1 stalk directs tRNA movements during translation
    Jingyi Fei
    Department of Chemistry, Columbia University, New York, NY 10027, USA
    Proc Natl Acad Sci U S A 106:15702-7. 2009
    ..Collectively, our data demonstrate that EF-G and the L1 stalk allosterically collaborate to direct tRNA translocation from the P to the E sites, and suggest a model for the release of E-site tRNA...
  89. pmc Single-molecule analysis of protein-free U2-U6 snRNAs
    Zhuojun Guo
    Department of Chemistry, Wayne State University, Detroit, Michigan, USA
    Nat Struct Mol Biol 16:1154-9. 2009
    ..Site-specific mutations in the highly conserved AGC triad and the U80 base in U6 suggest that the observed conformational dynamics correlate with residues that have an important role in splicing...
  90. pmc Shot-noise limited single-molecule FRET histograms: comparison between theory and experiments
    Eyal Nir
    Department of Chemistry and Biochemistry, and California NanoScience Institute, University of California at Los Angeles, Los Angeles, California 90095, USA
    J Phys Chem B 110:22103-24. 2006
    ..6 A. Finally, we briefly mention the results of a future publication and illustrate them with a simple two-state model system (DNA hairpin), for which the kinetic transition rates between the open and closed conformations are extracted...
  91. ncbi Protein biosensors based on the principle of fluorescence resonance energy transfer for monitoring cellular dynamics
    Isaac T Li
    Institute of Biomaterials and Biomedical Engineering, University of Toronto, 164 College Street, Toronto, Ont, Canada, M5S 3G9
    Biotechnol Lett 28:1971-82. 2006
    Genetically-coded, fluorescence resonance energy transfer (FRET) biosensors are widely used to study molecular events from single cells to whole organisms...
  92. ncbi FRET in cell biology: still shining in the age of super-resolution?
    Hernan E Grecco
    Department of Systemic Cell Biology, Max Planck Institute of Molecular Physiology, Otto Hahn Strasse 11, D 44227 Dortmund, Germany
    Chemphyschem 12:484-90. 2011
    ..Herein we review the major applications in biological FRET imaging and we discuss the possibilities and challenges in the super-resolution era...
  93. ncbi How Förster resonance energy transfer imaging improves the understanding of protein interaction networks in cancer biology
    Gilbert O Fruhwirth
    Richard Dimbleby Department of Cancer Research, Division of Cancer Studies, King s College London, Guy s Medical School Campus, NHH, SE1 1UL, UK
    Chemphyschem 12:442-61. 2011
    ....
  94. ncbi Graphene fluorescence resonance energy transfer aptasensor for the thrombin detection
    Haixin Chang
    Department of Chemistry, Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology, Tsinghua University, Beijing 100084, People s Republic of China
    Anal Chem 82:2341-6. 2010
    ..In this work, a highly sensitive and specific fluorescence resonance energy transfer (FRET) aptasensor for thrombin detection is developed based on the dye labeled aptamer assembled ..
  95. pmc FLIM FRET technology for drug discovery: automated multiwell-plate high-content analysis, multiplexed readouts and application in situ
    Sunil Kumar
    Photonics Group, Department of Physics, Imperial College London, London SW7 2AZ, UK
    Chemphyschem 12:609-26. 2011
    ..For cell-based assays, an exemplar application reading out protein aggregation using FLIM FRET is presented, and the potential for multiple simultaneous FLIM (FRET) readouts in microscopy is illustrated...
  96. pmc FRET microscopy in 2010: the legacy of Theodor Förster on the 100th anniversary of his birth
    Yuansheng Sun
    W M Keck Center for Cellular Imaging KCCI, Department of Biology, University of Virginia, Gilmer Hall 064, McCormick Rd, Charlottesville, VA 22904, USA
    Chemphyschem 12:462-74. 2011
    ....
  97. ncbi Single-molecule four-color FRET visualizes energy-transfer paths on DNA origami
    Ingo H Stein
    Angewandte Physik Biophysik, Amalienstrasse 54, Ludwig Maximilians Universitat, 80799 Munich, Germany
    J Am Chem Soc 133:4193-5. 2011
    b>Fluorescence resonance energy transfer (FRET) represents a mechanism to transport light energy at the nanoscale, as exemplified by nature's light-harvesting complexes...
  98. pmc Development of FRET assay into quantitative and high-throughput screening technology platforms for protein-protein interactions
    Yang Song
    Department of Bioengineering, Bourns College of Engineering, University of California at Riverside, 900 University Avenue, Riverside, CA 92521, USA
    Ann Biomed Eng 39:1224-34. 2011
    ..The FRET-based HTS is pioneer in cell-based HTS. Both K (d) determination and cell-based HTS, carried out in 384-well plate format, provide powerful tools for large-scale and high-throughput applications...
  99. pmc Visualizing dynamic cytoplasmic forces with a compliance-matched FRET sensor
    Fanjie Meng
    Center for Single Molecule Biophysics, Department of Physiology and Biophysics, State University of New York at Buffalo, 3435 Main Street, Buffalo, NY 14214, USA
    J Cell Sci 124:261-9. 2011
    ..Protein stress sensors are opening new perspectives in cell biology...
  100. pmc Self-illuminating in vivo lymphatic imaging using a bioluminescence resonance energy transfer quantum dot nano-particle
    Nobuyuki Kosaka
    Molecular Imaging Program, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD 20892 1088, USA
    Contrast Media Mol Imaging 6:55-9. 2011
    ..These advantages of BRET-QDot allowed us to perform real-time, quantitative lymphatic imaging without image processing. BRET-Qdots have the potential to be a robust nano-material platform for developing optical molecular imaging probes...
  101. ncbi Biophysical characterization of the fluorescent protein voltage probe VSFP2.3 based on the voltage-sensing domain of Ci-VSP
    Alicia Lundby
    Laboratory for Neuronal Circuit Dynamics, RIKEN Brain Science Institute, Wako City, Saitama, 351 0198, Japan
    Eur Biophys J 39:1625-35. 2010
    ..3. VSFP2.3 is a fluorescent voltage probe that reports changes in membrane potential as a FRET (fluorescence resonance energy transfer) signal. Here we report sensing current measurements from VSFP2.3, and show that VSFP2.3 carries 1...

Research Grants77

  1. High Sensitivity Multi-Well Plate Reader Based on an Array of Photon Counting Det
    William Lawrence; Fiscal Year: 2007
    ..detection methods include absorbance, fluorescence intensity and polarization, chemiluminescence, fluorescence resonance energy transfer (FRET) and time resolved fluorescence...
  2. Tissue-specific delivery of probes by control of membrane trafficking of endoprot
    Blake Peterson; Fiscal Year: 2010
    ..By flanking members of the peptide library with red and green fluorophores, fluorescence resonance energy transfer (FRET) will allow quantification of in vivo cleavage of members of the peptide library...
  3. Characterizing TLR Function of Human Dendritic Cells in the Elderly
    Alexander Panda; Fiscal Year: 2012
    ..PI will employ cell biology techniques including pulse-chase experiments, confocal microscopy and fluorescence resonance energy transfer to evaluate age-associated changes in the kinetics of TLR1 surface expression, intracellular ..
  4. Ionic currents in gastrointestinal smooth muscle
    Hamid I Akbarali; Fiscal Year: 2013
    ..We will also investigate the interaction of src kinase with the calcium channel using fluorescence resonance energy transfer (FRET) and siRNA approaches to delineate the functional effect of src-Ca2+ channel interaction in ..
  5. Post-translational regulation of MLL in leukemogenesis
    ANDREW GEORGE MUNTEAN; Fiscal Year: 2012
    ..MLL-PAFc binding will be monitored by fluorescence resonance energy transfer (FRET) and interaction of candidate chemical compounds will be verified by NMR structural analysis...
  6. Post-translational regulation of MLL in leukemogenesis
    ANDREW GEORGE MUNTEAN; Fiscal Year: 2013
    ..MLL-PAFc binding will be monitored by fluorescence resonance energy transfer (FRET) and interaction of candidate chemical compounds will be verified by NMR structural analysis...
  7. Mechanism and functional role of AKAP79/150 in M current control and excitability
    Mark S Shapiro; Fiscal Year: 2010
    ..Techniques to be used include fluorescence resonance energy transfer (FRET), total internal reflection fluorescence (TIRF), confocal microscopy, patch-clamp ..
  8. STRUCTURE AND FUNCTION OF PAXILLIN
    Christopher E Turner; Fiscal Year: 2012
    ..time-lapse microscopy, combined with Fluorescence Recovery after Photobleaching (FRAP) and Fluorescence Resonance Energy Transfer (FRET) analysis to evaluate cell morphology, polarity and migration as well focal adhesion ..
  9. MOLECULAR MODELS OF IMMUNE-MEDIATED VASCULAR INJURY
    Jeffrey R Bender; Fiscal Year: 2012
    ..of the T cell LFA-1-induced interactions between HuR and hnRNP-C, using pull-down methods, fluorescence resonance energy transfer (FRET) assays, and RNAi-based approaches;and (3) definitively evaluate the role of HuR in ..
  10. MOLECULAR PROPERTIES OF IONIC PERMEABILITY IN NERVE
    Bertil Hille; Fiscal Year: 2013
    ..The methodology will involve single-cell biophysics, electrophysiology, fluorescence resonance energy transfer, photometry, molecular biology, and confocal microscopy...
  11. FRET-based Biosensors to Monitor Redox in Cell Cycle Regulation
    Paul J A Kenis; Fiscal Year: 2012
    ..These constructs use fluorescence resonance energy transfer (FRET), a distance- and orientation- dependent energy transfer process between donor and acceptor ..
  12. Reciprocal Control of Ca2+ Channels by Anchored Protein Kinase A and Calcineurin
    William A Sather; Fiscal Year: 2010
    ..We plan to obtain from single living cells on a microscope stage dynamic measurements of fluorescence resonance energy transfer (FRET) between the various partners in this signaling complex, in an effort to uncover intra- and ..
  13. STRUCTURE AND FUNCTION OF NUCLEIC ACIDS
    Ignacio Tinoco; Fiscal Year: 2013
    ..to obtain this knowledge, newly discovered single-molecule methods are applied: Laser Tweezers and Fluorescence Resonance Energy Transfer (FRET). RNA molecules are synthesized in the laboratory by RNA polymerase from a DNA template...
  14. MRP4 extrudes cAMP for localized regulation of calcium channel activity
    OLIVIA RUTH BUONARATI; Fiscal Year: 2013
    ..Levels of cAMP will be monitored by Fluorescence Resonance Energy Transfer upon [unreadable]2AR stimulation with and without MRP4 inhibition or shRNA knockdown (KD)...
  15. Flow-induced coronary vasospasm in diabetic patients
    Zsolt Bagi; Fiscal Year: 2013
    ..to detect spatial distribution and interaction of Pecam-1 laser scanning confocal microscopy and fluorescence resonance energy transfer approaches will be used in isolated, pressurized coronary arteries and coronary endothelial cells ..
  16. LIPID MODULATION OF RHODOPSIN SIGNALING IN MEMBRANES
    Michael F Brown; Fiscal Year: 2010
    ..In addition, Fourier transform infrared (FTIR), fluorescence resonance energy transfer (FRET), and plasmon waveguide resonance (PWR) spectroscopy will elucidate the retinal environment, ..
  17. Adiponetin and AMPK Biosensor
    John Y J Shyy; Fiscal Year: 2010
    ..In addition, we developed a prototype of a fluorescence resonance energy transfer (FRET)-based AMPK activity reporter (AMPKAR) to measure AMPK activity...
  18. EBP50 Regulation of PTH Receptor in Bone
    Peter A Friedman; Fiscal Year: 2013
    ..to predict the effect of amino acid mutation on binding affinity, and biophysical measurements of fluorescence resonance energy transfer microscopy to acquire dynamic interactions in living cells and in real time, and surface plasmon ..
  19. FKBP Interaction with RYR Calcium Channels in Heart
    Donald M Bers; Fiscal Year: 2013
    ..relevance in cardiac myocytes and isolated SR and RyRs (using fluorescent- tagged FKBP and fluorescence resonance energy transfer (FRET). Four Specific Aims will: 1) identify key FKBP12 &12...
  20. NEUTROPHIL FLAVOCYTOCHROME B STRUCTURE AND FUNCTION
    ALGIRDAS JOSEPH JESAITIS; Fiscal Year: 2010
    ..to covalently modify these antibodies with fluorescent probes, to triangulate heme sites using fluorescence resonance energy transfer, measure the distance between antibody sites, and determine how these distances change upon ..
  21. Single Molecule Dynamics of mRNA Translation
    Yale E Goldman; Fiscal Year: 2013
    ..approach coupling Total Internal Reflection Fluorescence Microscopy (TIRFM) with single molecule Fluorescence Resonance Energy Transfer (smFRET)...
  22. Structural and Mechanistic Characterization of the Histone Chaperone FACT
    Duane David Winkler; Fiscal Year: 2012
    ..Second, solution-based binding, competition, and fluorescence resonance energy transfer (FRET) assays will provide important mechanistic information on FACT mediated nucleosome assembly/..
  23. Zeiss 2-photon (2P) LSM780 laser scanning confocal microscope
    Moshe Levi; Fiscal Year: 2013
    ....
  24. Design and Implementation of Genetically Encoded Myosin Based Force Sensors
    DANIEL PETER KIEHART; Fiscal Year: 2013
    ..Specific Aim 1: We will construct genetically encoded Tension Sensing Modules (TSMs) based on Fluorescence Resonance Energy Transfer (FRET) pairs separated by protein springs...
  25. Structural Dynamics of Cardiac Muscle Contraction
    Brett A Colson; Fiscal Year: 2012
    ..3) Use time-resolved phosphorescence anisotropy (TPA) and fluorescence resonance energy transfer (FRET), with probes on actin or myosin in solution, to determine the effects of purified ..
  26. Role of nAChR stoichiometry in nicotine-induced upregulation
    CHRISTOPHER I RICHARDS; Fiscal Year: 2012
    ..and cultured neurons will be investigated using total internal reflection microscopy (TIRFM) and fluorescence resonance energy transfer (FRET) at both the single vesicle and single receptor level...
  27. EGF Receptor Signaling in Time and Space in Tumor Cells
    Alexander D Sorkin; Fiscal Year: 2012
    ..using a combination of RNA interference, live-cell fluorescence imaging, multi-chromophore fluorescence resonance energy transfer microscopy and biochemical techniques, the mechanisms of the formation of the multi-protein ..
  28. Nano-sized Cell Guidance System for Ischemic Tissue Repair
    Hyunjoon Kong; Fiscal Year: 2010
    ..Specifically, we will use a fluorescence resonance energy transfer (FRET) technique we previously developed to quantify the number of poly(glycerol) bound to EPCs...
  29. STRUCTURE-FUNCTION RELATIONSHIPS OF IMMUNORECEPTORS
    Barbara A Baird; Fiscal Year: 2013
    ..Changes in proximity between Fc5RI and order- vs disorder- preferring lipids will be measured with fluorescence resonance energy transfer. Cholesterol distribution across the outer and inner leaflets of the plasma membrane will be ..
  30. MOLECULAR PROPERTIES OF IONIC PERMEABILITY IN NERVE
    Bertil Hille; Fiscal Year: 2012
    ..The methods will include patch-clamp electrophysiology, fluorescence resonance energy transfer, dynamic targeting of enzymes to cellular membranes, confocal microscopy, and chemical analysis...
  31. HIV integrase/DNA complexes and concerted integration
    Duane P Grandgenett; Fiscal Year: 2010
    ..In- gel fluorescence resonance energy transfer-derived distance measurements and atomic force microscopy will determine the topology of the two ..
  32. Novel Fluorescent Nucleotide Analogues to Probe Nucleic Acid Metabolism
    BYRON WILLIS PURSE; Fiscal Year: 2010
    ..expansion of the absorption and emission window and the brightening of the dyes, new FRET (Fluorescence Resonance Energy Transfer) applications and direct in vivo fluorescence imaging of DNA synthesis are likely to be possible...
  33. Homogenous HTS Assays to Screen for Inhibitors of Keap1-Nrf2 Interaction
    Longqin Hu; Fiscal Year: 2010
    ..and validated in this new R01 application using fluorescence polarization (FP) and time-resolved fluorescence resonance energy transfer (TR-FRET)...
  34. FRET Imaging of Molecular Hierarchy at Subcellular Levels in Mechanotransduction
    Yingxiao Wang; Fiscal Year: 2013
    ..b>Fluorescence resonance energy transfer (FRET) technology and genetically encoded biosensors have provided powerful tools for visualizing ..
  35. Spliceosome Mechanism Dissected at the Single Molecule Level
    Nils G Walter; Fiscal Year: 2013
    ..To address this challenge, we recently have developed single molecule fluorescence resonance energy transfer (smFRET) assays that have begun to dissect pre-mRNA conformational changes during splicing...
  36. Antigen Binding, Crosslinking and Transmembrane Signaling
    David A Holowka; Fiscal Year: 2013
    ..Ca2+ influx via recently identified Orai1/CRACM1 calcium channels will be characterized using fluorescence resonance energy transfer imaging for measurements of molecular proximity between this channel and the calcium sensor, STIM1...
  37. Site-Directed Oxidative Modification of Muscle Protein Structural Dynamics
    David D Thomas; Fiscal Year: 2013
    ..A complementary array of spectroscopic techniques will be used - fluorescence resonance energy transfer (FRET), transient phosphorescence anisotropy (TPA), electron paramagnetic resonance (EPR), and ..
  38. Signaling complex remodeling on the surfaces of live macrophages
    SHALINI THERESE LOW-NAM; Fiscal Year: 2013
    ..Clustering dynamics will be imaged using fluorescent chimeric receptor and quantified by Fluorescence Resonance Energy Transfer (FRET)...
  39. Spinning Disk Confocoal Microscope
    Judy Drazba; Fiscal Year: 2010
    ..It will also provide the ability to do fluorescence resonance energy transfer (FRET) measurements, fluorescence recovery after photobleaching (FRAP) measurements and ..
  40. HTS to identify chemical probes of the apelin receptor signaling pathway
    Layton Harris Smith; Fiscal Year: 2010
    ..The second assay uses the commercially available Lance homogeneous time-resolved fluorescence resonance energy transfer (TR-FRET) assay to measure cAMP accumulation...
  41. New probes for matrix metalloproteinase 13
    Gregg B Fields; Fiscal Year: 2013
    ..Our laboratory has developed triple-helical peptide (THP) fluorescence resonance energy transfer (FRET) substrates for high throughput screening (HTS) of collagenolytic MMPs and identification of ..
  42. Protein Kinase Regulatory Subunit I Directed Mutagenesis
    Susan S Taylor; Fiscal Year: 2013
    ..such as small angle Xray scattering, small angle neutron scattering, stopped-flow fluorescence, fluorescence resonance energy transfer, hydrogen/deuterium exchange coupled with mass spectrometry, and NMR spectroscopy in parallel with ..
  43. ION GRADIENTS AND ENERGY COUPLING IN BACTERIA
    Peter C Maloney; Fiscal Year: 2012
    ..work to place pairs of fluorescent probes at strategic positions on OxlT and use single-pair fluorescence resonance energy transfer (spFRET) to measure separation distances changes at the single-molecule level...
  44. Dynamics of nucleosome assembly/disassembly affected by chromatin modifications
    Tae Hee Lee; Fiscal Year: 2013
    ..In order to accomplish these aims, we will employ single molecule multi-color fluorescence resonance energy transfer to monitor the real-time dynamics of DNA wrapping/unwrapping during nucleosome assembly/..
  45. ApoE4 Structure Correctors as a Therapeutic Approach for Alzheimers Disease
    Robert W Mahley; Fiscal Year: 2013
    ..A secondary assay based on GFP- apoE4-eDHFR fluorescence resonance energy transfer will be used to confirm hits selectively...
  46. TSH RECEPTOR MULTIMERIZATION
    TERRY FRANCIS DAVIES; Fiscal Year: 2013
    ..more recently we modeled such forms in transfected cells by using functionally tagged TSHRs and fluorescence resonance energy transfer (FRET)...
  47. Analysis of Z-band assembly and maintenance in living skeletal muscle cells
    JOSEPH WILLIAM SANGER; Fiscal Year: 2013
    ..Steady-state FRET (Fluorescence Resonance Energy Transfer) efficiencies of the proteins and their binding partners co-expressed in living skeletal muscle ..
  48. Structural Basis of Dysfunctional Calcium Release Channels in Heart Disease
    Zheng Liu; Fiscal Year: 2013
    ..combine two highly complementary biophysical techniques, cryo- electron microscopy (cryo-EM) and fluorescence resonance energy transfer (FRET), to characterize functional structures and conformational dynamics of RyR2...
  49. Development of novel single-molecule approaches for nanoscale study of helicases
    Sanford H Leuba; Fiscal Year: 2010
    ..In this study, we will develop novel uses of single-pair Fluorescence Resonance Energy Transfer (spFRET), Magnetic Tweezers (MT) and combination instruments for the analysis of DNA-structure ..
  50. Large-Scale Conformational Changes in Regulation of Transcription Initiation
    M Thomas Record; Fiscal Year: 2011
    ..in concert with low resolution structural characterizations (chemical and enzymatic footprinting;fluorescence resonance energy transfer (FRET)) to determine what conformational changes occur in each step, and how each sets the stage ..
  51. Processing of Alternative Structures in Telomeric DNA
    GERALD JOSEPH NORA; Fiscal Year: 2011
    ..G4 DNA formation and dissociation will be tracked using single molecule fluorescence resonance energy transfer (FRET)...
  52. Mood-Stabilizing Medications and the Inositol Signaling System
    Ranga N Parthasarathy; Fiscal Year: 2013
    ..receptor-mediated endocytosis will be monitored by following transferrin receptor trafficking and fluorescence resonance energy transfer (FRET) analysis...
  53. Biophysics Cell Membrane Probe
    William E Brownell; Fiscal Year: 2010
    ..user specific experiments such as calcium imaging, fluorescence recovery after photobleaching or fluorescence resonance energy transfer may be easily accommodated...
  54. Activation and Signaling Mechanism of the RET Tyrosine Kinase Receptor
    JENNIFER MARIE CHOW; Fiscal Year: 2013
    ..of RET activation and signaling by pursuing following three aims: 1) genetically encoded fluorescence resonance energy transfer (FRET) constructs will be used to develop a reporter system for monitoring RET association events ..
  55. Single-molecule studies of ATP-dependent chromatin remodeling
    Xiaowei Zhuang; Fiscal Year: 2013
    ..In this project, we will use single-molecule fluorescence resonance energy transfer, in conjunction with complementary biochemical assays, to study the mechanisms of chromatin ..
  56. ZEISS LSM710 SCANNING CONFOCAL MICROSCOPE
    Hazel L Sive; Fiscal Year: 2010
    ..Lifetime Imaging (FLIM), which enables study of molecular lifetime, and facilitates FRET (Fluorescence Resonance Energy Transfer) analyses, to determine molecular interactions...
  57. Intramolecular domain movements of PcrA during its helicase function
    Karen R Thickman; Fiscal Year: 2010
    ..Using single-pair fluorescence resonance energy transfer (spFRET), magnetic tweezers and rotor beads, I propose to correlate the conformational changes ..
  58. Role of Cardiac Troponin in Health and Disease
    JONATHAN PAUL DAVIS; Fiscal Year: 2013
    ..We have developed a novel competition assay and a fluorescence resonance energy transfer (FRET) system that can probe the critical TnC-TnI interactions...
  59. Generation of an ITK Biosensor Tool Box
    Constantine D Tsoukas; Fiscal Year: 2010
    ..These constructs will be expressed in T cells and by using Fluorescence Resonance Energy Transfer (FRET) analysis we will assess whether ITK occurs in an intramolecular fold or an intermolecular ..
  60. Assay Development for Substrate and Phosphorylation State Specific JNK Inhibitors
    Philip LoGrasso; Fiscal Year: 2013
    ..FP) assay for discovering substrate and bidentate competitive selective inhibitors, and fluorescence resonance energy transfer (FRET) assays for activation (phosphorylation) state selective inhibitors...
  61. SENSORY TRANSDUCTION IN BACTERIAL CHEMOTAXIS
    Howard C Berg; Fiscal Year: 2013
    ..Using fluorescence resonance energy transfer (FRET) between fluorescent fusion proteins in vivo, we will study responses of cells to time-..
  62. Imaging Transporter Protein Dynamics in Living Cells
    TIM CHIFONG LEI; Fiscal Year: 2013
    ..raster image correlation spectroscopy (ccRICS), number &brightness (N&B) analysis, fluorescence resonance energy transfer (FRET), lifetime imaging, and molecular tracking (MT) techniques...
  63. Phospholemman and Na-Pump Function in Cardiac Myocytes
    Donald M Bers; Fiscal Year: 2013
    ..that PLM physically associates with NKA 1PLM phosphorylation (via immunoprecipitation), but in fluorescence resonance energy transfer (FRET) measurements, that PLM phosphorylation alters the PLM-NKA interaction and that PLM forms ..
  64. PLATELET SUFACE ACTIVATION IN HEMOSTASIS AND THROMBOSIS
    SANFORD SHATTIL; Fiscal Year: 1993
    ..In parallel studies, fluorescence resonance energy transfer techniques will be used to examine whether clustering of receptors is required for these ..
  65. Regulation of Epithelial Sodium Channel by CFTR chloride channel
    Bakhrom Berdiev; Fiscal Year: 2009
    ..We will determine the CFTR-ENaC interaction using Fluorescence Resonance Energy Transfer (FRET) microscopy and co-immunoprecipitation (co-IP)...
  66. Novel Extended Red-Shifted Rhodamine Dyes
    NICHOLAS CAIRNS; Fiscal Year: 2004
    ..that are designed for use in high throughput screening (HTS) fluorescent polarization (FP) and fluorescence resonance energy transfer (FRET) biological assays for new drug and agrochemical discovery research...
  67. DNA Sequencing at a Stretch
    Di Gao; Fiscal Year: 2009
    ..Through a fluorescence resonance energy transfer (FRET) mechanism, detachment of the DNA strand can be directly detected from the fluorescence ..
  68. Single Molecule Protein and Biopolymer Folding Dynamics
    Shimon Weiss; Fiscal Year: 2003
    ..We will use single-pair fluorescence resonance energy transfer (spFRET) as reporter of the distance between two amino acid residues in the polypeptide chain, or ..
  69. Trafficking of BMP signals in neurons
    Guillermo Marques; Fiscal Year: 2007
    ..A combination of Bimolecular Fluorescence Complementation, (BiFC), Fluorescence Resonance Energy Transfer (FRET) and live imaging of the fluorescently tagged components of the pathway (ligands, receptors ..
  70. PRE & POSTSYNAPTIC NICOTINIC ACHRS--STRUCTURE & FUNCTION
    Peter Sargent; Fiscal Year: 1999
    ..Finally, at the molecular level, we will use fluorophore-tagged antibodies and fluorescence resonance energy transfer (FRET) to examine the proximity of subunits to each other...
  71. Discovery of novel fluorescent reporter genes
    Ming Zhao; Fiscal Year: 2004
    ..special importance for minimizing background emission and scattering in vivo as well as for FRET (fluorescence resonance energy transfer) analysis...
  72. Real-time measurement of ethanol's effect on cyclic AMP metabolism in live cells
    Masami Yoshimura; Fiscal Year: 2009
    ..polypeptide chain cAMP sensor molecules have been developed, which can monitor cAMP levels by fluorescence resonance energy transfer (FRET)...
  73. Spectroscopy of myosin light chain domain dynamics
    Osha Roopnarine; Fiscal Year: 2010
    ..using two complementary spectroscopic techniques: electron paramagnetic resonance (EPR) and fluorescence resonance energy transfer (FRET)...
  74. Lipid modulation of beta-cell calcium channels
    Lina Moitoso de Vargas; Fiscal Year: 2006
    ..Our preliminary data from the fluorescence resonance energy transfer analyses indicate that the L-type channel predominantly expressed in beta-cells, the ..
  75. Real-time Single-Molecule Analysis for Transient Molec. Complex Characterization
    Taiho Kim; Fiscal Year: 2009
    DESCRIPTION (provided by applicant): Single-molecule fluorescence resonance energy transfer (smFRET) has rapidly developed to answer fundamental questions about biological mechanisms, providing detailed views of molecular machines at ..
  76. Hybrid Nanostructures for Wireless In Vivo Action Potential Sensing
    Joshua Goldberger; Fiscal Year: 2009
    ..The specific hypothesis behind the proposed research is that semiconductor nanoparticle Fluorescence Resonance Energy Transfer (FRET) dyes which emit and are excited in the near IR spectrum, can enhance the signal to noise of ..
  77. A Novel HTS Cell-Based Assay For Cyclic GMP
    Gary Piazza; Fiscal Year: 2007
    ..b>Fluorescence resonance energy transfer (FRET) results from a conformational change of the protein upon cGMP binding to cause decreased ..