Rotational diffusion of the α(2a) adrenergic receptor revealed by FlAsH labeling in living cells.


:The fluorescein arsenical hairpin binder (FlAsH) shows much promise to determine the relative orientations of protein regions and structures even in living cells and in the plasma membrane. In this study, we characterized FlAsH's photophysical properties by steady-state anisotropy and time-resolved single photon counting for further applications with G-protein coupled receptors. We find that FlAsH has a relatively high initial anisotropy of 0.31 ± 0.01 and a three-component fluorescence lifetime with an average of 4.1 ± 0.1 ns. We characterized the FlAsH fluorophore orientation in the α(2A) adrenergic receptor revealing rigid orientations of FlAsH in the membrane plane for rotational correlation times of ∼50 ns in living cells. To elucidate the fluorophore-membrane orientation and rotational correlation time, an anisotropy treatment similar to that of another researcher (Axelrod, D. 1979. Biophys. J. 26:557-573) was developed. The rotational correlation times were observed to increase by up to 16 ns after agonist addition. The rotational correlation time also allowed for a comparison to the theoretical relationship between translational and rotational diffusion (originally proposed by Saffman, P. G., and M. Delbrück. 1975. Proc. Natl. Acad. Sci. USA. 72:3111-3113) and revealed a discrepancy of a factor between 10 and 100.


Biophys J


Biophysical journal


Spille JH,Zürn A,Hoffmann C,Lohse MJ,Harms GS




Has Abstract


2011-02-16 00:00:00














  • Sorting of lipids and transmembrane peptides between detergent-soluble bilayers and detergent-resistant rafts.

    abstract::Specific proteins and lipids sequester to regions of cell membranes called rafts. Due to their high content of sphingomyelin (SM) and cholesterol, raft bilayers are thicker than nonraft bilayers and, at least at 4 degrees C, are resistant to Triton X-100 extraction. It has been postulated that rafts concentrate protei...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: McIntosh TJ,Vidal A,Simon SA

    更新日期:2003-09-01 00:00:00

  • Flexibility of ras lipid modifications studied by 2H solid-state NMR and molecular dynamics simulations.

    abstract::Human posttranslationally modified N-ras oncogenes are known to be implicated in numerous human cancers. Here, we applied a combination of experimental and computational techniques to determine structural and dynamical details of the lipid chain modifications of an N-ras heptapeptide in 1,2-dimyristoyl-sn-glycero-3-ph...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Vogel A,Tan KT,Waldmann H,Feller SE,Brown MF,Huster D

    更新日期:2007-10-15 00:00:00

  • Compartmentalization of the outer hair cell demonstrated by slow diffusion in the extracisternal space.

    abstract::In the outer hair cell (OHC), the extracisternal space (ECiS) is a conduit and reservoir of the molecular and ionic substrates of the lateral wall, including those necessary for electromotility. To determine the mechanisms through which molecules are transported in the ECiS of the OHC, we selectively imaged the time-d...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Gliko O,Saggau P,Brownell WE

    更新日期:2009-08-19 00:00:00

  • Energy transfer and charge separation kinetics in photosystem I: Part 1: Picosecond transient absorption and fluorescence study of cyanobacterial photosystem I particles.

    abstract::The energy transfer and charge separation kinetics of a photosystem I (PS I) core particle of an antenna size of 100 chlorophyll/P700 has been studied by combined fluorescence and transient absorption kinetics with picosecond resolution. This is the first combined picosecond study of transient absorption and fluoresce...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Holzwarth AR,Schatz G,Brock H,Bittersmann E

    更新日期:1993-06-01 00:00:00

  • Configurational entropy change of netropsin and distamycin upon DNA minor-groove binding.

    abstract::Binding of a small molecule to a macromolecular target reduces its conformational freedom, resulting in a negative entropy change that opposes the binding. The goal of this study is to estimate the configurational entropy change of two minor-groove-binding ligands, netropsin and distamycin, upon binding to the DNA dup...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Dolenc J,Baron R,Oostenbrink C,Koller J,van Gunsteren WF

    更新日期:2006-08-15 00:00:00

  • Self-assembly and conformational heterogeneity of the AXH domain of ataxin-1: an unusual example of a chameleon fold.

    abstract::Ataxin-1 is a human protein responsible for spinocerebellar ataxia type 1, a hereditary disease associated with protein aggregation and misfolding. Essential for ataxin-1 aggregation is the anomalous expansion of a polyglutamine tract near the protein N-terminus, but the sequence-wise distant AXH domain modulates and ...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: de Chiara C,Rees M,Menon RP,Pauwels K,Lawrence C,Konarev PV,Svergun DI,Martin SR,Chen YW,Pastore A

    更新日期:2013-03-19 00:00:00

  • Structural and functional similarities between the nucleotide-binding domains of CFTR and GTP-binding proteins.

    abstract::The opening and closing of the CFTR Cl- channel are regulated by ATP hydrolysis at its two nucleotide binding domains (NBDs). However, the mechanism and functional significance of ATP hydrolysis are unknown. Sequence similarity between the NBDs of CFTR and GTP-binding proteins suggested the NBDs might have a structure...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Carson MR,Welsh MJ

    更新日期:1995-12-01 00:00:00

  • Protein logic: a statistical mechanical study of signal integration at the single-molecule level.

    abstract::Information processing and decision-making is based upon logic operations, which in cellular networks has been well characterized at the level of transcription. In recent years, however, both experimentalists and theorists have begun to appreciate that cellular decision-making can also be performed at the level of a s...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: de Ronde W,Rein ten Wolde P,Mugler A

    更新日期:2012-09-05 00:00:00

  • Conformational Dynamics of the HIV-Vif Protein Complex.

    abstract::Human immunodeficiency virus-1 viral infectivity factor (Vif) is an intrinsically disordered protein responsible for the ubiquitination of the APOBEC3 (A3) antiviral proteins. Vif folds when it binds Cullin-RING E3 ligase 5 and the transcription cofactor CBF-β. A five-protein complex containing the substrate receptor ...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Ball KA,Chan LM,Stanley DJ,Tierney E,Thapa S,Ta HM,Burton L,Binning JM,Jacobson MP,Gross JD

    更新日期:2019-04-23 00:00:00

  • Concerted gating mechanism underlying KATP channel inhibition by ATP.

    abstract::KATP channels assemble from four regulatory SUR1 and four pore-forming Kir6.2 subunits. At the single-channel current level, ATP-dependent gating transitions between the active burst and the inactive interburst conformations underlie inhibition of the KATP channel by intracellular ATP. Previously, we identified a slow...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Drain P,Geng X,Li L

    更新日期:2004-04-01 00:00:00

  • The inhibition of bacteriophage DNA injection by dyes bound to the DNA.

    abstract::A delay ( approximately 10 min) in the appearance of intracellular phage is caused by preincubating the infecting phage T4o(1) in proflavine, acridine orange, or ethidium, but not polyamines. No significant delay in attachment is observed. Apparently the presence of the dye is required inside the permeability barrier ...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Dusenbery DB,Uretz RB

    更新日期:1972-08-01 00:00:00

  • The intrinsic pKa values for phosphatidylserine and phosphatidylethanolamine in phosphatidylcholine host bilayers.

    abstract::Potentiometric titrations and surface potential measurements have been used to determine the intrinsic pKa values of both the carboxyl and amino groups of phosphatidylserine (PS) in mixed vesicles of PS and phosphatidylcholine (PC), and also of the amino group of phosphatidylethanolamine (PE) in mixed PE-PC vesicles. ...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Tsui FC,Ojcius DM,Hubbell WL

    更新日期:1986-02-01 00:00:00

  • Effects of conformational peptide probe DP4 on bidirectional signaling between DHPR and RyR1 calcium channels in voltage-clamped skeletal muscle fibers.

    abstract::In skeletal muscle, excitation-contraction coupling involves the activation of dihydropyridine receptors (DHPR) and type-1 ryanodine receptors (RyR1) to produce depolarization-dependent sarcoplasmic reticulum Ca²⁺ release via orthograde signaling. Another form of DHPR-RyR1 communication is retrograde signaling, in whi...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Olojo RO,Hernández-Ochoa EO,Ikemoto N,Schneider MF

    更新日期:2011-05-18 00:00:00

  • DNA multiphoton absorption generates localized damage for studying repair dynamics in live cells.

    abstract::Investigations into the spatiotemporal dynamics of DNA repair using live-cell imaging are aided by the ability to generate well defined regions of ultravioletlike photolesions in an optical microscope. We demonstrate that multiphoton excitation of DNA in live cells with visible femtosecond pulses produces thymine cycl...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Daddysman MK,Fecko CJ

    更新日期:2011-11-02 00:00:00

  • The conduction of protons in different stereoisomers of dioxolane-linked gramicidin A channels.

    abstract::Two different stereoisomers of the dioxolane-linked gramicidin A (gA) channels were individually synthesized (the SS and RR dimers;. Science. 244:813-817). The structural differences between these dimers arise from different chiralities within the dioxolane linker. The SS dimer mimics the helicity and the inter- and i...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Quigley EP,Quigley P,Crumrine DS,Cukierman S

    更新日期:1999-11-01 00:00:00

  • Instabilities in the transient response of muscle.

    abstract::We investigate the isometric transient response of muscle using a quantitative stochastic model of the actomyosin cycle based on the swinging lever-arm hypothesis. We first consider a single pair of filaments, and show that when values of parameters such as the lever-arm displacement and the cross-bridge elasticity ar...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Vilfan A,Duke T

    更新日期:2003-08-01 00:00:00

  • Ligand-receptor kinetics measured by total internal reflection with fluorescence correlation spectroscopy.

    abstract::Total internal reflection excitation used in combination with fluorescence correlation spectroscopy (TIR-FCS) is a method for characterizing the dynamic behavior and absolute concentrations of fluorescent molecules near or at the interface of a planar substrate and a solution. In this work, we demonstrate for the firs...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Lieto AM,Cush RC,Thompson NL

    更新日期:2003-11-01 00:00:00

  • The conserved phenylalanine in the K+ channel voltage-sensor domain creates a barrier with unidirectional effects.

    abstract::Voltage-gated ion channels are crucial for regulation of electric activity of excitable tissues such as nerve cells, and play important roles in many diseases. During activation, the charged S4 segment in the voltage sensor domain translates across a hydrophobic core forming a barrier for the gating charges. This barr...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Schwaiger CS,Liin SI,Elinder F,Lindahl E

    更新日期:2013-01-08 00:00:00

  • The Affinity of Sterols for Different Phospholipid Classes and Its Impact on Lateral Segregation.

    abstract::Cholesterol is an essential molecule in the membranes of mammalian cells. It is known to be distributed heterogeneously within the cells, between the bilayer leaflets, as well as between lateral domains within the bilayer. However, we do not know exactly how cholesterol is distributed and what forces drive this sortin...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Nyholm TKM,Jaikishan S,Engberg O,Hautala V,Slotte JP

    更新日期:2019-01-22 00:00:00

  • Tissue-specific mathematical models of slow wave entrainment in wild-type and 5-HT(2B) knockout mice with altered interstitial cells of Cajal networks.

    abstract::Gastrointestinal slow waves are generated within networks of interstitial cells of Cajal (ICCs). In the intact tissue, slow waves are entrained to neighboring ICCs with higher intrinsic frequencies, leading to active propagation of slow waves. Degradation of ICC networks in humans is associated with motility disorders...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Du P,O'Grady G,Gibbons SJ,Yassi R,Lees-Green R,Farrugia G,Cheng LK,Pullan AJ

    更新日期:2010-05-19 00:00:00

  • Testing of aggregation measurement techniques for intramembranous particles.

    abstract::Under various physiological and nonphysiological conditions, the intramembranous particles, as seen by freeze-fracture electron microscopy, may be in various degrees of aggregation. To compare various schemes for the measurement of the degree of aggregation, a computer program has been used to generate simulated aggre...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Donnell JT 2nd,Finegold LX

    更新日期:1981-09-01 00:00:00

  • Spontaneous-curvature theory of clathrin-coated membranes.

    abstract::Clathrin-coated membranes are precursors to coated vesicles in the receptor-mediated endocytic pathway. In this paper we present a physical model for the first steps of the transformation of a clathrin-coated membrane into a coated vesicle. The theory is based on in vitro cytoplasmic acidification experiments of Heuse...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Mashl RJ,Bruinsma RF

    更新日期:1998-06-01 00:00:00

  • Termination of cardiac Ca(2+) sparks: an investigative mathematical model of calcium-induced calcium release.

    abstract::A Ca(2+) spark arises when a cluster of sarcoplasmic reticulum (SR) channels (ryanodine receptors or RyRs) opens to release calcium in a locally regenerative manner. Normally triggered by Ca(2+) influx across the sarcolemmal or transverse tubule membrane neighboring the cluster, the Ca(2+) spark has been shown to be t...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Sobie EA,Dilly KW,dos Santos Cruz J,Lederer WJ,Jafri MS

    更新日期:2002-07-01 00:00:00

  • Physical model for self-organization of actin cytoskeleton and adhesion complexes at the cell front.

    abstract::Cell motion is driven by interplay between the actin cytoskeleton and the cell adhesions in the front part of the cell. The actin network segregates into lamellipodium and lamellum, whereas the adhesion complexes are characteristically distributed underneath the actin system. Here, we suggest a computational model for...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Shemesh T,Bershadsky AD,Kozlov MM

    更新日期:2012-04-18 00:00:00

  • Change of motion and localization of cholesterol molecule during L(alpha)-H(II) transition.

    abstract::Formation of the inverted hexagonal (H(II)) phase from the lamellar (L(alpha)) phase of bovine brain-extracted phosphatidylcholine (BBPC) and phosphatidylethanolamine (BBPE) was investigated using 31P-NMR with or without cholesterol. When the ratio of BBPC to BBPE was 1:1, the H(II) formation was observed in the prese...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Hayakawa E,Naganuma M,Mukasa K,Shimozawa T,Araiso T

    更新日期:1998-02-01 00:00:00

  • Pulsatile signaling in intercellular communication. Periodic stimuli are more efficient than random or chaotic signals in a model based on receptor desensitization.

    abstract::The efficiency of various patterns of pulsatile stimulation is determined in a model in which a receptor becomes desensitized in the presence of its stimulatory ligand. The effect of stochastic or chaotic changes in the duration and/or interval between successive pulses in a series of square-wave stimuli is investigat...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Li Y,Goldbeter A

    更新日期:1992-01-01 00:00:00

  • Spectroscopic and molecular modeling studies of caffeine complexes with DNA intercalators.

    abstract::Recent studies have demonstrated that caffeine can act as an antimutagen and inhibit the cytoxic and/or cytostatic effects of some DNA intercalating agents. It has been suggested that this inhibitory effect may be due to complexation of the DNA intercalator with caffeine. In this study we employ optical absorption, fl...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Larsen RW,Jasuja R,Hetzler RK,Muraoka PT,Andrada VG,Jameson DM

    更新日期:1996-01-01 00:00:00

  • A nuclear magnetic resonance investigation of the energetics of basepair opening pathways in DNA.

    abstract::The opening of basepairs plays a key role in DNA replication and transcription, and in the action of DNA repair and modification enzymes. In this article, we have used proton exchange to define the energetics of the pathways for basepair opening in two DNA 17-mer duplexes. The rates of exchange of imino protons with s...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Coman D,Russu IM

    更新日期:2005-11-01 00:00:00

  • Deuterium-Enhanced Raman Spectroscopy for Histidine pKa Determination in a pH-Responsive Hydrogel.

    abstract::We report here a method for the determination of the pKa of histidine in complex or heterogeneous systems amenable to neither solid-state nor solution NMR spectroscopy. Careful synthesis of a fluorenylmethyloxycarbonyl- and trityl-protected, C2-deuterated histidine produces a vibrational-probe-equipped amino acid that...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Braun GA,Pogostin BH,Pucetaite M,Londergan CH,Åkerfeldt KS

    更新日期:2020-11-03 00:00:00

  • The Berg-Purcell limit revisited.

    abstract::Biological systems often have to measure extremely low concentrations of chemicals with high precision. When dealing with such small numbers of molecules, the inevitable randomness of physical transport processes and binding reactions will limit the precision with which measurements can be made. An important question ...

    journal_title:Biophysical journal

    pub_type: 杂志文章


    authors: Kaizu K,de Ronde W,Paijmans J,Takahashi K,Tostevin F,ten Wolde PR

    更新日期:2014-02-18 00:00:00