A cellular screening assay using analysis of metal-modified fluorescence lifetime.


:Current methods for screening cell receptor internalization often require complex image analysis with limited sensitivity. Here we describe a novel bioassay based on detection of changes in global fluorescence lifetime above a gold substrate, with superresolution axial sensitivity and no need for image analysis. We show that the lifetime of enhanced green fluorescent protein expressed in a cellular membrane is greatly reduced in close proximity to the gold, resulting in a distance-dependent lifetime distribution throughout the cell. We demonstrate the application of this phenomenon in a screening assay by comparing the efficacies of two small molecule inhibitors interfering with the internalization process of a G protein-coupled receptor.


Biophys J


Biophysical journal


Cade NI,Fruhwirth G,Archibald SJ,Ng T,Richards D




Has Abstract


2010-06-02 00:00:00














  • Association thermodynamics and conformational stability of beta-sheet amyloid beta(17-42) oligomers: effects of E22Q (Dutch) mutation and charge neutralization.

    abstract::Amyloid fibrils are associated with many neurodegenerative diseases. It was found that amyloidogenic oligomers, not mature fibrils, are neurotoxic agents related to these diseases. Molecular mechanisms of infectivity, pathways of aggregation, and molecular structure of these oligomers remain elusive. Here, we use all-...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Blinov N,Dorosh L,Wishart D,Kovalenko A

    更新日期:2010-01-20 00:00:00

  • Modulation of 4-AP block of a mammalian A-type K channel clone by channel gating and membrane voltage.

    abstract::We examined the state-, voltage-, and time dependences of interaction between 4-AP and a mammalian A-type K channel clone (rKv1.4) expressed in Xenopus oocytes using whole-cell and single-channel recordings. 4-AP blocked rKv1.4 from the cytoplasmic side of the membrane. The development of block required channel openin...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Yao JA,Tseng GN

    更新日期:1994-07-01 00:00:00

  • Quantitative analysis of G-actin transport in motile cells.

    abstract::Cell migration is based on an actin treadmill, which in turn depends on recycling of G-actin across the cell, from the rear where F-actin disassembles, to the front, where F-actin polymerizes. To analyze the rates of the actin transport, we used the Virtual Cell software to solve the diffusion-drift-reaction equations...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Novak IL,Slepchenko BM,Mogilner A

    更新日期:2008-08-01 00:00:00

  • Trauma of the erythrocyte membrane associated with low shear stress.

    abstract::Studies have been performed on erythrocytes that have been subjected to a low shear stress of less than 100 dyn/cm(2) in a cone-and-plate viscometer. Alterations that were observed included decreased red cell survival, increased osmotic fragility, changes in the cation permeability of the red cell membrane, and a redu...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Nanjappa BN,Chang HK,Glomski CA

    更新日期:1973-11-01 00:00:00

  • Kinetics of microtubule catastrophe assessed by probabilistic analysis.

    abstract::Microtubules are cytoskeletal filaments whose self-assembly occurs by abrupt switching between states of roughly constant growth and shrinkage, a process known as dynamic instability. Understanding the mechanism of dynamic instability offers potential for controlling microtubule-dependent cellular processes such as ne...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Odde DJ,Cassimeris L,Buettner HM

    更新日期:1995-09-01 00:00:00

  • Voltage-dependent removal of sodium inactivation by N-bromoacetamide and pronase.

    abstract::When perfused internally through crayfish giant axons, pronase removed sodium inactivation more than three times as fast at -100 mV as compared with -30 mV. N-bromoacetamide, applied internally, removed sodium inactivation twice as fast at -100 mV as at -30 mV, and the relative rate of removal declined with membrane d...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Salgado VL,Yeh JZ,Narahashi T

    更新日期:1985-04-01 00:00:00

  • Mechanics of sperm-egg interaction at the zona pellucida.

    abstract::Mammalian sperm traverse several layers of egg vestments before fertilization can occur. The innermost vestment, the zona pellucida, is a glycoprotein shell, which captures and tethers the sperm before they penetrate it. We report here direct measurements of the force required to tether a motile human sperm as well as...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Baltz JM,Katz DF,Cone RA

    更新日期:1988-10-01 00:00:00

  • A systematic methodology for defining coarse-grained sites in large biomolecules.

    abstract::Coarse-grained (CG) models of biomolecules have recently attracted considerable interest because they enable the simulation of complex biological systems on length-scales and timescales that are inaccessible for atomistic molecular dynamics simulation. A CG model is defined by a map that transforms an atomically detai...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Zhang Z,Lu L,Noid WG,Krishna V,Pfaendtner J,Voth GA

    更新日期:2008-12-01 00:00:00

  • Transition states in protein folding kinetics: modeling phi-values of small beta-sheet proteins.

    abstract::Small single-domain proteins often exhibit only a single free-energy barrier, or transition state, between the denatured and the native state. The folding kinetics of these proteins is usually explored via mutational analysis. A central question is which structural information on the transition state can be derived fr...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Weikl TR

    更新日期:2008-02-01 00:00:00

  • Role of constraint in catalysis and high-affinity binding by proteins.

    abstract::Using a model for catalysis of a dynamic equilibrium, the role of constraint in catalysis is quantified. The intrinsic rigidity of proteins is shown to be insufficient to constrain the activated complexes of enzymes, irrespective of the mechanism. However, when minimization of the surface excess free energy of water s...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Vanselow DG

    更新日期:2002-05-01 00:00:00

  • The mechanism of inactivation of T4 bacteriophage by tritium decay.

    abstract::Coliphage T4 was used as a model system to study the mechanism of biological inactivation produced by tritium decay. Experimentally, tritiated precursors were incorporated into phage DNA (thymidine-(3)H) or into phage protein ((3)H-amino acids). The ratio of killing efficiencies for decays originating in phage DNA to ...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Funk F,Person S,Bockrath RC Jr

    更新日期:1968-09-01 00:00:00

  • Backstepping Mechanism of Kinesin-1.

    abstract::Kinesin-1 is an ATP-driven molecular motor that transports cellular cargo along microtubules. At low loads, kinesin-1 almost always steps forward, toward microtubule plus ends, but at higher loads, it can also step backward. Backsteps are usually 8 nm but can be larger. These larger backward events of 16 nm, 24 nm, or...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Toleikis A,Carter NJ,Cross RA

    更新日期:2020-11-17 00:00:00

  • Non-Newtonian behavior of blood in oscillatory flow.

    abstract::Sinusoidal oscillatory flow of blood and of aqueous glycerol solutions was produced in rigid cylindrical tubes. For aqueous glycerol, the amplitude of the measured pressure gradient wave form conformed closely to that predicted by Womersley's theory of oscillatory flow, up to Reynolds numbers approaching 2000. Blood d...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Kunz AL,Coulter NA Jr

    更新日期:1967-01-01 00:00:00


    abstract::Fission fragment tracks from uranium-stained biological preparations have been observed in the electron microscope in a study of the feasibility of their use as tracers. Virus, DNA, and tissue sections were stained with uranium, prepared for electron microscopy, irradiated in the high thermal neutron flux of a nuclear...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: MALMON AG

    更新日期:1964-01-01 00:00:00

  • Modeling Endoplasmic Reticulum Network Maintenance in a Plant Cell.

    abstract::The endoplasmic reticulum (ER) in plant cells forms a highly dynamic network of complex geometry. ER network morphology and dynamics are influenced by a number of biophysical processes, including filament/tubule tension, viscous forces, Brownian diffusion, and interactions with many other organelles and cytoskeletal e...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Lin C,White RR,Sparkes I,Ashwin P

    更新日期:2017-07-11 00:00:00

  • Low-frequency stimulation induces stable transitions in stereotypical activity in cortical networks.

    abstract::Reverberating spontaneous synchronized brain activity is believed to play an important role in neural information processing. Whether and how external stimuli can influence this spontaneous activity is poorly understood. Because periodic synchronized network activity is also prominent in in vitro neuronal cultures, we...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Vajda I,van Pelt J,Wolters P,Chiappalone M,Martinoia S,van Someren E,van Ooyen A

    更新日期:2008-06-01 00:00:00

  • An electron spin resonance study of synaptosome opiate receptors. The preparation and use of a spin labeled morphine.

    abstract::Morphine spin labeled on the phenolic hydroxy group has been prepared using commercially available reagents and characterized by thin layer chromatography, mass spectroscopy, and electron spin resonance spectroscopy. It has been shown that morphine modified in this way retains some opiate activity, does not pass throu...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Copeland ES,Boykin ME,Kelley JA,Kullberg MP

    更新日期:1975-11-01 00:00:00

  • Voltage-dependent block by saxitoxin of sodium channels incorporated into planar lipid bilayers.

    abstract::We have previously studied single, voltage-dependent, saxitoxin-(STX) blockable sodium channels from rat brain in planar lipid bilayers, and found that channel block by STX was voltage-dependent. Here we describe the effect of voltage on the degree of block and on the kinetics of the blocking reaction. From their volt...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: French RJ,Worley JF 3rd,Krueger BK

    更新日期:1984-01-01 00:00:00

  • Spectroscopy on the B850 band of individual light-harvesting 2 complexes of Rhodopseudomonas acidophila. I. Experiments and Monte Carlo simulations.

    abstract::The electronic structure of the circular aggregate of 18 bacteriochlorophyll a (BChl a) molecules responsible for the B850 absorption band of the light-harvesting 2 (LH2) complex of the photosynthetic purple bacterium Rhodopseudomonas acidophila has been studied by measuring fluorescence-excitation spectra of individu...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Ketelaars M,van Oijen AM,Matsushita M,Köhler J,Schmidt J,Aartsma TJ

    更新日期:2001-03-01 00:00:00

  • Phosphorus-31 two-dimensional solid-state exchange NMR. Application to model membrane and biological systems.

    abstract::Two-dimensional solid-state 31P NMR has been used to investigate the orientational exchange of phospholipids in gel and liquid-crystalline aqueous multilamellar dispersions and oriented multibilayers, and in biological membranes. In liquid-crystalline L alpha multilamellar dispersions, orientational exchange originate...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Fenske DB,Jarrell HC

    更新日期:1991-01-01 00:00:00

  • Comparison of the dynamics of the primary events of bacteriorhodopsin in its trimeric and monomeric states.

    abstract::In this paper, femtosecond pump-probe spectroscopy in the visible region of the spectrum has been used to examine the ultrafast dynamics of the retinal excited state in both the native trimeric state and the monomeric state of bacteriorhodopsin (bR). It is found that the excited state lifetime (probed at 490 nm) incre...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Wang J,Link S,Heyes CD,El-Sayed MA

    更新日期:2002-09-01 00:00:00

  • Protein self-association in solution: the bovine pancreatic trypsin inhibitor decamer.

    abstract::We have used magnetic relaxation dispersion to study bovine pancreatic trypsin inhibitor (BPTI) self-association as a function of pH, salt type and concentration, and temperature. The magnetic relaxation dispersion method sensitively detects stable oligomers without being affected by other interactions. We find that B...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Gottschalk M,Venu K,Halle B

    更新日期:2003-06-01 00:00:00

  • A delayed rectifier potassium current in Xenopus oocytes.

    abstract::A delayed voltage-dependent K+ current endogenous to Xenopus oocytes has been investigated by the voltage-clamp technique. Both activation and inactivation of the K+ current are voltage-dependent processes. The K+ currents were activated when membrane potential was depolarized from a holding potential of -90 to -50 mV...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Lu L,Montrose-Rafizadeh C,Hwang TC,Guggino WB

    更新日期:1990-06-01 00:00:00

  • Thermodynamics and kinetics of a folded-folded' transition at valine-9 of a GCN4-like leucine zipper.

    abstract::Spin inversion transfer (SIT) NMR experiments are reported probing the thermodynamics and kinetics of interconversion of two folded forms of a GCN4-like leucine zipper near room temperature. The peptide is 13Calpha-labeled at position V9(a) and results are compared with prior findings for position L13(e). The SIT data...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: d'Avignon DA,Bretthorst GL,Holtzer ME,Holtzer A

    更新日期:1999-05-01 00:00:00

  • Ultrafast absorption difference spectra of the Fenna-Matthews-Olson protein at 19 K: experiment and simulations.

    abstract::We describe simulations of absorption difference spectra in strongly coupled photosynthetic antennas. In the presence of large resonance couplings, distinctive features arise from excited-state absorption transitions between one- and two-exciton levels. We first outline the theory for the heterodimer and for the gener...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Buck DR,Savikhin S,Struve WS

    更新日期:1997-01-01 00:00:00

  • The nonequilibrium phase and glass transition behavior of beta-lactoglobulin.

    abstract::Concentrated solutions of bovine beta-lactoglobulin were studied using osmotic stress and rheological techniques. At pH 6.0 and 8.0, the osmotic pressure was largely independent of NaCl concentration and could be described by a hard sphere equation of state. At pH 5.1, close to the isoelectric point, the osmotic press...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Parker R,Noel TR,Brownsey GJ,Laos K,Ring SG

    更新日期:2005-08-01 00:00:00

  • Evolution of the hemifused intermediate on the pathway to membrane fusion.

    abstract::The pathway to membrane fusion in synthetic and biological systems is thought to pass through hemifusion, in which the outer leaflets are fused while the inner leaflets engage in a hemifusion diaphragm (HD). Fusion has been proposed to be completed by lysis of the expanded HD that matures from a localized stalklike in...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Warner JM,O'Shaughnessy B

    更新日期:2012-08-22 00:00:00

  • More gating charges are needed to open a Shaker K+ channel than are needed to open an rBIIA Na+ channel.

    abstract::This study presents what is, to our knowledge, a novel technique by means of which the ratio of the single gating charges of voltage-gated rat brain IIA (rBIIA) sodium and Shaker potassium ion channels was estimated. In the experiment, multiple tandems of enhanced green fluorescent protein were constructed and inserte...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Gamal El-Din TM,Grögler D,Lehmann C,Heldstab H,Greeff NG

    更新日期:2008-08-01 00:00:00

  • Minor groove deformability of DNA: a molecular dynamics free energy simulation study.

    abstract::The conformational deformability of nucleic acids can influence their function and recognition by proteins. A class of DNA binding proteins including the TATA box binding protein binds to the DNA minor groove, resulting in an opening of the minor groove and DNA bending toward the major groove. Explicit solvent molecul...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Zacharias M

    更新日期:2006-08-01 00:00:00

  • Analytical calculation of intracellular calcium wave characteristics.

    abstract::We present a theoretical analysis of intracellular calcium waves propagated by calcium feedback at the inositol 1,4,5-trisphosphate (IP3) receptor. The model includes essential features of calcium excitability, but is still analytically tractable. Formulas are derived for the wave speed, amplitude, and width. The calc...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Kupferman R,Mitra PP,Hohenberg PC,Wang SS

    更新日期:1997-06-01 00:00:00