Doubly Spin-Labeled RNA as an EPR Reporter for Studying Multicomponent Supramolecular Assemblies.

Abstract:

:mRNAs are involved in complicated supramolecular complexes with human 40S and 80S ribosomes responsible for the protein synthesis. In this work, a derivative of nonaribonucleotide pUUCGUAAAA with nitroxide spin labels attached to the 5'-phosphate and to the C8 atom of the adenosine in sixth position (mRNA analog) was used for studying such complexes using double electron-electron resonance/pulsed electron-electron double resonance spectroscopy. The complexes were assembled with participation of tRNA(Phe), which targeted triplet UUC of the derivative to the ribosomal peptidyl site and predetermined location of the adjacent GUA triplet coding for Val at the aminoacyl (A) site. The interspin distances were measured between the two labels of mRNA analog attached to the first nucleotide of the peptidyl site bound codon and to the third nucleotide of the A site bound codon, in the absence/presence of second tRNA bound at the A site. The values of the obtained interspin distances agree with those calculated for available near-atomic structures of similar complexes of 40S and 80S ribosomes, showing that neither 60S subunit nor tRNA at the A site have a noticeable effect on arrangement of mRNA at the codon-anticodon interaction area. In addition, the shapes of distance distributions in four studied ribosomal complexes allowed conclusions on conformational flexibility of mRNA in these complexes. Overall, the results of this study are the first, to our knowledge, demonstration of double electron-electron resonance/pulsed electron-electron double resonance application for measurements of intramolecular distances in multicomponent supramolecular complexes involving intricate cellular machineries and for evaluating dynamic properties of ligands bound to these machineries.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Malygin AA,Graifer DM,Meschaninova MI,Venyaminova AG,Krumkacheva OA,Fedin MV,Karpova GG,Bagryanskaya EG

doi

10.1016/j.bpj.2015.10.042

subject

Has Abstract

pub_date

2015-12-15 00:00:00

pages

2637-2643

issue

12

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(15)01119-4

journal_volume

109

pub_type

杂志文章
  • Na/Ca exchange and Na/K-ATPase function are equally concentrated in transverse tubules of rat ventricular myocytes.

    abstract::Formamide-induced detubulation of rat ventricular myocytes was used to investigate the functional distribution of the Na/Ca exchanger (NCX) and Na/K-ATPase between the t-tubules and external sarcolemma. Detubulation resulted in a 32% decrease in cell capacitance, whereas cell volume was unchanged. Thus, the surface-to...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(03)74758-4

    authors: Despa S,Brette F,Orchard CH,Bers DM

    更新日期:2003-11-01 00:00:00

  • THE NUMERICAL SOLUTION OF THE TIME-DEPENDENT NERNST-PLANCK EQUATIONS.

    abstract::Calculations are reported of the time-dependent Nernst-Planck equations for a thin permeable membrane between electrolytic solutions. Charge neutrality is assumed for the time-dependent case. The response of such a membrane system to step current input is measured in terms of the time and space changes in concentratio...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/s0006-3495(65)86707-8

    authors: COHEN H,COOLEY JW

    更新日期:1965-03-01 00:00:00

  • An analysis of the membrane potential along a clamped squid axon.

    abstract::A partially depolarized squid axon membrane is assumed to have a quasi-steady state negative resistance, the membrane potential is clamped at one point, and a distribution of potential along the axon is obtained from the cable equation. Nominal experimental values of -2 ohm cm(2) for the membrane and 6 ohm cm(2) for t...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/s0006-3495(61)86898-7

    authors: COLE KS

    更新日期:1961-05-01 00:00:00

  • Spot variation fluorescence correlation spectroscopy allows for superresolution chronoscopy of confinement times in membranes.

    abstract::Resolving the dynamical interplay of proteins and lipids in the live-cell plasma membrane represents a central goal in current cell biology. Superresolution concepts have introduced a means of capturing spatial heterogeneity at a nanoscopic length scale. Similar concepts for detecting dynamical transitions (superresol...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2011.04.035

    authors: Ruprecht V,Wieser S,Marguet D,Schütz GJ

    更新日期:2011-06-08 00:00:00

  • Comparative Study of the Condensing Effects of Ergosterol and Cholesterol.

    abstract::Cholesterol, due to its condensing effect, is considered an important regulator of membrane thickness. Other sterols, due to their structural similarities to cholesterol, are often assumed to have a universal effect on membrane properties similar to the condensing effect of cholesterol, albeit possibly to different de...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2016.04.003

    authors: Hung WC,Lee MT,Chung H,Sun YT,Chen H,Charron NE,Huang HW

    更新日期:2016-05-10 00:00:00

  • Fluorescence correlation microscopy of cells in the presence of autofluorescence.

    abstract::Fluorescence correlation microscopy (FCM), the combination of fluorescence correlation spectroscopy (FCS) and digital microscopy (Brock and Jovin, 1998. Cell. Mol. Biol. 44:847-856), has been implemented for measuring molecular diffusion and association in living cells with explicit consideration of autocorrelations a...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(98)77699-4

    authors: Brock R,Hink MA,Jovin TM

    更新日期:1998-11-01 00:00:00

  • Interaction of toposome from sea-urchin yolk granules with dimyristoyl phosphatidylserine model membranes: a 2H-NMR study.

    abstract::The yolk granule is the most abundant membrane-bound organelle present in sea urchin eggs and embryos. The major protein component of this organelle, toposome, accounts for approximately 50% of the total yolk protein and has been shown to be localized to the embryonic cell surface. Extensive characterization in severa...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.106.089979

    authors: Hayley M,Emberley J,Davis PJ,Morrow MR,Robinson JJ

    更新日期:2006-12-15 00:00:00

  • Testing for microscopic reversibility in the gating of maxi K+ channels using two-dimensional dwell-time distributions.

    abstract::An assumption usually made when developing kinetic models for the gating of ion channels is that the transitions among the various states involved in the gating obey microscopic reversibility. If this assumption is incorrect, then the models and estimated rate constants made with the assumption would be in error. This...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(94)80458-8

    authors: Song L,Magleby KL

    更新日期:1994-07-01 00:00:00

  • Raman microscope and quantum yield studies on the primary photochemistry of A2-visual pigments.

    abstract::The 77-K resonance Raman vibrational spectrum of intact goldfish rod photoreceptors containing 3,4-dehydro (A2) retinal is dominated by scattering from the 9-cis component of the steady state at all excitation wavelengths. Intact goldfish photoreceptors were regenerated with an A1-retinal chromophore to determine whet...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(87)83250-2

    authors: Barry B,Mathies RA,Pardoen JA,Lugtenburg J

    更新日期:1987-10-01 00:00:00

  • The enthalpy of acyl chain packing and the apparent water-accessible apolar surface area of phospholipids.

    abstract::The energetics of phospholipid aggregation depend on the apparent water-accessible apolar surface area (ASAap), ordering effects of the chains, and headgroup interactions. We quantify the enthalpy and entropy of these interactions separately. For that purpose, the thermodynamics of micelle formation of lysophosphatidy...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(01)76012-2

    authors: Heerklotz H,Epand RM

    更新日期:2001-01-01 00:00:00

  • Chemical denaturation: potential impact of undetected intermediates in the free energy of unfolding and m-values obtained from a two-state assumption.

    abstract::The chemical unfolding transition of a protein was simulated, including the presence of an intermediate (I) in equilibrium with the native (N) and unfolded (U) states. The calculations included free energies of unfolding, DeltaGuw, in the range of 1.4 kcal/mol to 10 kcal/mol and three different global m-values. The si...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(98)77537-X

    authors: Soulages JL

    更新日期:1998-07-01 00:00:00

  • A Model for the Hippo Pathway in the Drosophila Wing Disc.

    abstract::Although significant progress has been made toward understanding morphogen-mediated patterning in development, control of the size and shape of tissues via local and global signaling is poorly understood. In particular, little is known about how cell-cell interactions are involved in the control of tissue size. The Hi...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2018.07.002

    authors: Gou J,Lin L,Othmer HG

    更新日期:2018-08-21 00:00:00

  • Proton transport through influenza A virus M2 protein reconstituted in vesicles.

    abstract::Influenza A virus M2 protein is known to form acid-activated, proton-selective, amantadine-sensitive channels. We directly measured proton uptake in vesicles containing reconstituted M2 by monitoring external pH after addition of valinomycin to vesicles with 100-fold-diluted external [K(+)]. External pH typically incr...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.107.109082

    authors: Moffat JC,Vijayvergiya V,Gao PF,Cross TA,Woodbury DJ,Busath DD

    更新日期:2008-01-15 00:00:00

  • Binding of cations of group IA and IIA to bovine serum amine oxidase: effect on the activity.

    abstract::In this paper, we report on the presence of cation binding areas on bovine serum amine oxidase, where metal ions of the groups IA and IIA, such as Na(+), K(+), Cs(+), Mg(2+), and Ca(2+), bind with various affinities. We found a cation-binding area that influences the enzyme activity if occupied, so that the catalytic ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(02)73983-0

    authors: Di Paolo ML,Scarpa M,Corazza A,Stevanato R,Rigo A

    更新日期:2002-10-01 00:00:00

  • Arg(615)Cys substitution in pig skeletal ryanodine receptors increases activation of single channels by a segment of the skeletal DHPR II-III loop.

    abstract::The effect of peptides, corresponding to sequences in the skeletal muscle dihydropyridine receptor II-III loop, on Ca(2+) release from sarcoplasmic reticulum (SR) and on ryanodine receptor (RyR) calcium release channels have been compared in preparations from normal and malignant hyperthermia (MH)-susceptible pigs. Pe...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(01)76147-4

    authors: Gallant EM,Curtis S,Pace SM,Dulhunty AF

    更新日期:2001-04-01 00:00:00

  • A General Workflow for Characterization of Nernstian Dyes and Their Effects on Bacterial Physiology.

    abstract::The electrical membrane potential (Vm) is one of the components of the electrochemical potential of protons across the biological membrane (proton motive force), which powers many vital cellular processes. Because Vm also plays a role in signal transduction, measuring it is of great interest. Over the years, a variety...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2019.10.030

    authors: Mancini L,Terradot G,Tian T,Pu Y,Li Y,Lo CJ,Bai F,Pilizota T

    更新日期:2020-01-07 00:00:00

  • Activation-inactivation coupling in Myxicola giant axons injected with tetraethylammonium.

    abstract::Myxicola giant axons internally injected with tetraethylammonium chloride to block potassium currents were examined under voltage clamp. The sodium inactivation time constants obtained from the decline in INa during step depolarizations were substantially smaller than those obtained using conditioning prepulses to the...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(76)85749-9

    authors: Schauf CL,Pencek TL,Davis FA

    更新日期:1976-09-01 00:00:00

  • A multiscale red blood cell model with accurate mechanics, rheology, and dynamics.

    abstract::Red blood cells (RBCs) have highly deformable viscoelastic membranes exhibiting complex rheological response and rich hydrodynamic behavior governed by special elastic and bending properties and by the external/internal fluid and membrane viscosities. We present a multiscale RBC model that is able to predict RBC mecha...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2010.02.002

    authors: Fedosov DA,Caswell B,Karniadakis GE

    更新日期:2010-05-19 00:00:00

  • Force versus axial deflection of pipette-aspirated closed membranes.

    abstract::The axial deformation of a pipette-pressurized fluid membrane bag produces minuscule yet well-defined, reproducible forces. The stiffness of this ultrasensitive force transducer is tunable and largely independent of the constitutive membrane behavior. Based on a rigorous variational treatment, we present both numerica...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.107.104091

    authors: Heinrich V,Ounkomol C

    更新日期:2007-07-15 00:00:00

  • How many states can the motor molecule, prestin, assume in an electric field?

    abstract::By using an analogy between the magnetization of a paramagnetic material in an external magnetic field and the electric polarization of the lateral wall of outer hair cells in response to the transmembrane potential, we show that, based on experimental data on the charge transfer across the membrane, it is impossible ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.105.059782

    authors: Scherer MP,Gummer AW

    更新日期:2005-05-01 00:00:00

  • The role of diffusion in enzyme kinetics.

    abstract::A discussion is given of the general role of diffusion in enzyme kinetics based upon a rigorous theory for bimolecular association and dissociation steps that was presented previously, and criteria are formulated for the dependence of the over-all rate on medium viscosity. With these criteria it is possible to conclud...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(70)86331-7

    authors: Schurr JM

    更新日期:1970-08-01 00:00:00

  • Distributions in protein conformation space: implications for structure prediction and entropy.

    abstract::By considering how polymer structures are distributed in conformation space, we show that it is possible to quantify the difficulty of structural prediction and to provide a measure of progress for prediction calculations. The critical issue is the probability that a conformation is found within a specified distance o...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.104.041723

    authors: Sullivan DC,Kuntz ID

    更新日期:2004-07-01 00:00:00

  • An optical monitor of tension for small cardiac preparations.

    abstract::When a light beam is focused on a muscle preparation which is allowed to contract, large changes in the intensity of the emerging light accompany the contraction. These movement-related optical signals were studied and compared to simultaneous measurements of force in isolated cardiac Purkinje fibers. The two signals ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(81)84843-6

    authors: Kass RS

    更新日期:1981-04-01 00:00:00

  • Growing actin networks form lamellipodium and lamellum by self-assembly.

    abstract::Many different cell types are able to migrate by formation of a thin actin-based cytoskeletal extension. Recently, it became evident that this extension consists of two distinct substructures, designated lamellipodium and lamellum, which differ significantly in their kinetic and kinematic properties as well as their b...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.108.134817

    authors: Huber F,Käs J,Stuhrmann B

    更新日期:2008-12-15 00:00:00

  • Synthesis of voltage-sensitive fluorescence signals from three-dimensional myocardial activation patterns.

    abstract::Voltage-sensitive fluorescent dyes are commonly used to measure cardiac electrical activity. Recent studies indicate, however, that optical action potentials (OAPs) recorded from the myocardial surface originate from a widely distributed volume beneath the surface and may contain useful information regarding intramura...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(03)74690-6

    authors: Hyatt CJ,Mironov SF,Wellner M,Berenfeld O,Popp AK,Weitz DA,Jalife J,Pertsov AM

    更新日期:2003-10-01 00:00:00

  • Model inspired by nuclear pore complex suggests possible roles for nuclear transport receptors in determining its structure.

    abstract::Nuclear transport receptors (NTRs) mediate nucleocytoplasmic transport via their affinity for unstructured proteins (polymers) in the nuclear pore complex (NPC). Here, we have modeled the effect of NTRs on polymeric structure in the nanopore confinement of the NPC central conduit. The model explicitly takes into accou...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2013.11.013

    authors: Osmanović D,Ford IJ,Hoogenboom BW

    更新日期:2013-12-17 00:00:00

  • Effects of redox and sulfhydryl reagents on the bioelectric properties of the giant axon of the squid.

    abstract::The effects of internally and externally applied sulfhydryl reagents on the bioelectric properties of the giant axon of the squid Loligo pealeii and Dosidicus gigas were studied. Cysteine-HCl (400 mM, pH 7.3) was used to remove axoplasm from the perfusion channel. Oxidizing agents (1 to 60 mM) tended to increase the d...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(66)86686-9

    authors: Huneeus-Cox F,Fernandez HL,Smith BH

    更新日期:1966-09-01 00:00:00

  • Cholesterol level regulates endosome motility via Rab proteins.

    abstract::The role of cholesterol in the regulation of endosome motility was investigated by monitoring the intracellular trafficking of endocytosed folate receptors (FRs) labeled with fluorescent folate conjugates. Real-time fluorescence imaging of HeLa cells transfected with green fluorescent protein-tubulin revealed that FR-...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.106.099366

    authors: Chen H,Yang J,Low PS,Cheng JX

    更新日期:2008-02-15 00:00:00

  • High Spatiotemporal-Resolution Magnetic Tweezers: Calibration and Applications for DNA Dynamics.

    abstract::The observation of biological processes at the molecular scale in real time requires high spatial and temporal resolution. Magnetic tweezers are straightforward to implement, free of radiation or photodamage, and provide ample multiplexing capability, but their spatiotemporal resolution has lagged behind that of other...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2015.10.018

    authors: Dulin D,Cui TJ,Cnossen J,Docter MW,Lipfert J,Dekker NH

    更新日期:2015-11-17 00:00:00

  • Peptides derived from apoptotic Bax and Bid reproduce the poration activity of the parent full-length proteins.

    abstract::Bax and Bid are proapoptotic proteins of the Bcl-2 family that regulate the release of apoptogenic factors from mitochondria. Although they localize constitutively in the cytoplasm, their apoptotic function is exerted at the mitochondrial outer membrane, and is related to their ability to form transbilayer pores. Here...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.104.058008

    authors: García-Sáez AJ,Coraiola M,Dalla Serra M,Mingarro I,Menestrina G,Salgado J

    更新日期:2005-06-01 00:00:00