Force unfolding kinetics of RNA using optical tweezers. I. Effects of experimental variables on measured results.

Abstract:

:Experimental variables of optical tweezers instrumentation that affect RNA folding/unfolding kinetics were investigated. A model RNA hairpin, P5ab, was attached to two micron-sized beads through hybrid RNA/DNA handles; one bead was trapped by dual-beam lasers and the other was held by a micropipette. Several experimental variables were changed while measuring the unfolding/refolding kinetics, including handle lengths, trap stiffness, and modes of force applied to the molecule. In constant-force mode where the tension applied to the RNA was maintained through feedback control, the measured rate coefficients varied within 40% when the handle lengths were changed by 10-fold (1.1-10.2 Kbp); they increased by two- to threefold when the trap stiffness was lowered to one-third (from 0.1 to 0.035 pN/nm). In the passive mode, without feedback control and where the force applied to the RNA varied in response to the end-to-end distance change of the tether, the RNA hopped between a high-force folded-state and a low-force unfolded-state. In this mode, the rates increased up to twofold with longer handles or softer traps. Overall, the measured rates remained with the same order-of-magnitude over the wide range of conditions studied. In the companion article on pages 3010-3021, we analyze how the measured kinetics parameters differ from the intrinsic molecular rates of the RNA, and thus how to obtain the molecular rates.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Wen JD,Manosas M,Li PT,Smith SB,Bustamante C,Ritort F,Tinoco I Jr

doi

10.1529/biophysj.106.094052

subject

Has Abstract

pub_date

2007-05-01 00:00:00

pages

2996-3009

issue

9

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(07)71109-8

journal_volume

92

pub_type

杂志文章
  • Proline scan of the HERG channel S6 helix reveals the location of the intracellular pore gate.

    abstract::In Shaker-like channels, the activation gate is formed at the bundle crossing by the convergence of the inner S6 helices near a conserved proline-valine-proline motif, which introduces a kink that allows for electromechanical coupling with voltage sensor motions via the S4-S5 linker. Human ether-a-go-go-related gene (...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2014.01.035

    authors: Thouta S,Sokolov S,Abe Y,Clark SJ,Cheng YM,Claydon TW

    更新日期:2014-03-04 00:00:00

  • Theory for ligand rebinding at cell membrane surfaces.

    abstract::Conditions for which a ligand reversibly bound to a cell surface dissociates and then rebinds to the surface have been theoretically examined. The coupled differential equations that describe reaction at the interface between sites on a plane and three-dimensional solution have been described previously (Thompson, N. ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(98)77836-1

    authors: Lagerholm BC,Thompson NL

    更新日期:1998-03-01 00:00:00

  • Frequency domain analysis of asymmetry current in squid axon membrane.

    abstract::The change in capacity of squid axon membrane during hyper- and depolarizations was investigated in the absence of ionic currents after the membrane was treated with pronase. In the presence of the inactivation process (h parameter), failure to observe the gating current in the frequency domain was attributed to the r...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(78)85475-7

    authors: Takashima S

    更新日期:1978-04-01 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

  • Time course of activation of calcium release from sarcoplasmic reticulum in skeletal muscle.

    abstract::Myoplasmic free calcium transients were measured with antipyrylazo III in voltage clamped segments of frog skeletal muscle fibers and were used to calculate the rate of release (Rrel) of calcium from the sarcoplasmic reticulum. Intramembrane charge movement was measured for the same pulses in the same fibers. During a...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(88)83050-9

    authors: Simon BJ,Schneider MF

    更新日期:1988-12-01 00:00:00

  • Effect of fluorine substitution on the interaction of lipophilic ions with the plasma membrane of mammalian cells.

    abstract::The effects of the anionic tungsten carbonyl complex [W(CO)(5)SC(6)H(5)](-) and its fluorinated analog [W(CO)(5)SC(6)F(5)](-) on the electrical properties of the plasma membrane of mouse myeloma cells were studied by the single-cell electrorotation technique. At micromolar concentrations, both compounds gave rise to a...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(00)76400-9

    authors: Kürschner M,Nielsen K,von Langen JR,Schenk WA,Zimmermann U,Sukhorukov VL

    更新日期:2000-09-01 00:00:00

  • Low frequency sounds from sustained contraction of human skeletal muscle.

    abstract::Low frequency audible vibrations are produced by human skeletal muscles undergoing sustained contraction. The effect is easily demonstrable with an electronic stethoscope which amplifies sound below 50 Hz. Autocorrelation analysis of the signal shows that it is periodic with a frequency 25 +/- 2.5 Hz. The quality of t...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(80)85080-6

    authors: Oster G,Jaffe JS

    更新日期:1980-04-01 00:00:00

  • Kinetic evidence for three photolyzable taxonomic conformational substates in oxymyoglobin.

    abstract::The kinetics of oxygen geminate binding with the taxonomic substates of MbO2 are reported. The maximum entropy method was used to analyze the rebinding kinetics of MbCO and MbO2 monitored in the Soret. The resulting rate distributions were found to consist of a small number of overlapping bands. A global parametric fi...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(02)75561-6

    authors: Tetreau C,Novikov E,Tourbez M,Lavalette D

    更新日期:2002-04-01 00:00:00

  • Multiwavelength analysis of the kinetics of reduction of cytochrome aa3 by cytochrome c.

    abstract::Some new approaches to the kinetic study of the reduction of cytochrome aa3 by cytochrome c are presented. The primary innovations are the use of a spectrometer which can acquire multiwavelength data as fast as every 10 microseconds, and the application of a variety of analytical methods which can utilize simultaneous...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(93)81170-6

    authors: Hendler RW,Bose SK,Shrager RI

    更新日期:1993-09-01 00:00:00

  • The connection between chromatin motion on the 100 nm length scale and core histone dynamics in live XTC-2 cells and isolated nuclei.

    abstract::The diffusive motion of DNA-containing chromatin in live cells and isolated nuclei is investigated using a two-photon standing wave fluorescence photobleaching experiment with 100 nm spatial resolution. The chromatin is labeled using the minor groove binding dye Hoechst 33342. In live cells, the mean diffusion rate is...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(04)74134-X

    authors: Davis SK,Bardeen CJ

    更新日期:2004-01-01 00:00:00

  • Free energy potential for aggregation of giant, neutral lipid bilayer vesicles by Van der Waals attraction.

    abstract::Here, we report the first direct observation of Van der Waals' attraction between biomembrane capsules using measurements of the free energy reduction per unit area of membrane-membrane contact formation. In these studies, the membrane capsules were reconstituted neutral (egg phosphatidylcholine) lipid bilayers of gia...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(84)84039-4

    authors: Evans E,Metcalfe M

    更新日期:1984-09-01 00:00:00

  • Mg2+-paracrystal formation of tropomyosin as a condensation phenomenon. Effects of pH, salt, temperature, and troponin binding.

    abstract::Tropomyosin (Tm) paracrystal formation induced by Mg2+ was studied by monitoring increases in light scattering. Paracrystals formed above a critical Tm concentration with lag phases in the time courses at pH 7.5 and 6.0, indicating that condensation polymerization processes are involved. The kinetic data at pH 7.5 rea...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(89)82655-4

    authors: Ishii Y,Lehrer SS

    更新日期:1989-07-01 00:00:00

  • Temperature-Induced Catch-Slip to Slip Bond Transit in Plasmodium falciparum-Infected Erythrocytes.

    abstract::Plasmodium falciparum malaria-infected red blood cells (IRBCs), or erythrocytes, avoid splenic clearance by adhering to host endothelium. Upregulation of endothelial receptors intercellular adhesion molecule-1 (ICAM-1) and cluster of differentiation 36 (CD36) are associated with severe disease pathology. Most in vitro...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2019.11.016

    authors: Lim YB,Thingna J,Kong F,Dao M,Cao J,Lim CT

    更新日期:2020-01-07 00:00:00

  • Detection of jumps in single-channel data containing subconductance levels.

    abstract::Detection algorithms are widely used for the analysis of single-channel data because they remove the background noise from the measured current signal and reconstruct the noise-free time series. Standard detection algorithms assume channels switching only between zero and full conductance. Many types of channels, howe...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(94)80614-9

    authors: Draber S,Schultze R

    更新日期:1994-10-01 00:00:00

  • Fluorescence fluctuation spectroscopy in the presence of immobile fluorophores.

    abstract::Fluorescence contributions from immobile sources present a challenge for fluorescence fluctuation spectroscopy (FFS) because the absence of signal fluctuations from stationary fluorophores leads to a biased analysis. This is especially of concern for cellular FFS studies on proteins that interact with immobile structu...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.107.115642

    authors: Skinner JP,Chen Y,Müller JD

    更新日期:2008-03-15 00:00:00

  • Global analysis of fluorescence lifetime imaging microscopy data.

    abstract::Global analysis techniques are described for frequency domain fluorescence lifetime imaging microscopy (FLIM) data. These algorithms exploit the prior knowledge that only a limited number of fluorescent molecule species whose lifetimes do not vary spatially are present in the sample. Two approaches to implementing the...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(00)76759-2

    authors: Verveer PJ,Squire A,Bastiaens PI

    更新日期:2000-04-01 00:00:00

  • Membrane surface-charge titration probed by gramicidin A channel conductance.

    abstract::We manipulate lipid bilayer surface charge and gauge its influence on gramicidin A channel conductance by two strategies: titration of the lipid charge through bulk solution pH and dilution of a charged lipid by neutral. Using diphytanoyl phosphatidylserine (PS) bilayers with CsCl aqueous solutions, we show that the e...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(98)77620-9

    authors: Rostovtseva TK,Aguilella VM,Vodyanoy I,Bezrukov SM,Parsegian VA

    更新日期:1998-10-01 00:00:00

  • Carboxyl tail prevents yeast K(+) channel closure: proposal of an integrated model of TOK1 gating.

    abstract::TOK1 encodes the channel responsible for the prominent outward K(+) current of the yeast plasma membrane. It can dwell in several impermeable states, including a rapidly transiting, K(+)-electromotive-force-dependent "R" (rectifying) state, a voltage-independent "IB" (interburst) state, and a set of [K(+)](ext) and vo...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(02)75440-4

    authors: Loukin SH,Saimi Y

    更新日期:2002-02-01 00:00:00

  • The beta-strand-loop-beta-strand conformation is marginally populated in beta2-microglobulin (20-41) peptide in solution as revealed by replica exchange molecular dynamics simulations.

    abstract::Solid-state NMR study shows that the 22-residue K3 peptide (Ser(20)-Lys(41)) from beta(2)-microglobulin (beta(2)m) adopts a beta-strand-loop-beta-strand conformation in its fibril state. Residue Pro(32) has a trans conformation in the fibril state of the peptide, while it adopts a cis conformation in the native state ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.107.125054

    authors: Liang C,Derreumaux P,Mousseau N,Wei G

    更新日期:2008-07-01 00:00:00

  • Variable symmetry in Salmonella typhimurium flagellar motors.

    abstract::Electron cryomicroscopy of rotor complexes of the Salmonella typhimurium flagellar motor, overproduced in a nonmotile Escherichia coli host, has revealed a variation in subunit symmetry of the cytoplasmic ring (C ring) module. C rings with subunit symmetries ranging from 31 to 38 were found. They formed a Gaussian dis...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(03)74877-2

    authors: Young HS,Dang H,Lai Y,DeRosier DJ,Khan S

    更新日期:2003-01-01 00:00:00

  • NMR studies in dodecylphosphocholine of a fragment containing the seventh transmembrane helix of a G-protein-coupled receptor from Saccharomyces cerevisiae.

    abstract::The structure and dynamics of a large segment of Ste2p, the G-protein-coupled alpha-factor receptor from yeast, were studied in dodecylphosphocholine (DPC) micelles using solution NMR spectroscopy. We investigated the 73-residue peptide EL3-TM7-CT40 consisting of the third extracellular loop 3 (EL3), the seventh trans...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.106.103770

    authors: Neumoin A,Arshava B,Becker J,Zerbe O,Naider F

    更新日期:2007-07-15 00:00:00

  • Phosphatidylethanolamine-phosphatidylglycerol bilayer as a model of the inner bacterial membrane.

    abstract::Phosphatidylethanolamine (PE) and phosphatidylglycerol (PG) are the main lipid components of the inner bacterial membrane. A computer model for such a membrane was built of palmitoyloleoyl PE (POPE) and palmitoyloleoyl PG (POPG) in the proportion 3:1, and sodium ions (Na+) to neutralize the net negative charge on each...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.104.048835

    authors: Murzyn K,Róg T,Pasenkiewicz-Gierula M

    更新日期:2005-02-01 00:00:00

  • The mechanical stability of immunoglobulin and fibronectin III domains in the muscle protein titin measured by atomic force microscopy.

    abstract::The domains of the giant muscle protein titin (connectin) provide interaction sites for other sarcomeric proteins and fulfill mechanical functions. In this paper we compare the unfolding forces of defined regions of different titin isoforms by single-molecule force spectroscopy. Constructs comprising six to eight immu...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(98)77741-0

    authors: Rief M,Gautel M,Schemmel A,Gaub HE

    更新日期:1998-12-01 00:00:00

  • TEM-1 backbone dynamics-insights from combined molecular dynamics and nuclear magnetic resonance.

    abstract::Dynamic properties of class A beta-lactamase TEM-1 are investigated from molecular dynamics (MD) simulations. Comparison of MD-derived order parameters with those obtained from model-free analysis of nuclear magnetic resonance (NMR) relaxation data shows high agreement for N-H moieties within alpha- and beta-secondary...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2009.08.061

    authors: Fisette O,Morin S,Savard PY,Lagüe P,Gagné SM

    更新日期:2010-02-17 00:00:00

  • Design Parameters for Granzyme-Mediated Cytotoxic Lymphocyte Target-Cell Killing and Specificity.

    abstract::Cytotoxic lymphocytes are key elements of the immune system that are primarily responsible for targeting cells infected with intracellular pathogens, or cells that have become malignantly transformed. Target cells are killed mainly via lymphocyte exocytosis of specialized lysosomes containing perforin, a pore-forming ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2015.06.045

    authors: Woodsworth DJ,Dunsing V,Coombs D

    更新日期:2015-08-04 00:00:00

  • Time-resolved absorption and magnetic circular dichroism spectroscopy of cytochrome c3 from Desulfovibrio.

    abstract::The UV-visible absorption and magnetic circular dichroism (MCD) spectra of the ferric, ferrous, CO-ligated forms and kinetic photolysis intermediates of the tetraheme electron-transfer protein cytochrome c3 (Cc3) are reported. Consistent with bis-histidinyl axial coordination of the hemes in this Class III c-type cyto...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(93)81226-8

    authors: O'Connor DB,Goldbeck RA,Hazzard JH,Kliger DS,Cusanovich MA

    更新日期:1993-10-01 00:00:00

  • The dependence of impulse propagation speed on firing frequency, dispersion, for the Hodgkin-Huxley model.

    abstract::Propagation speed of an impulse is influenced by previous activity. A pulse following its predecessor too closely may travel more slowly than a solitary pulse. In contrast, for some range of interspike intervals, a pulse may travel faster than normal because of a possible superexcitable phase of its predecessor's wake...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(81)84847-3

    authors: Miller RN,Rinzel J

    更新日期:1981-05-01 00:00:00

  • Observation of two families of folding pathways of BBL.

    abstract::BBL is an independent folding domain of a large multienzyme complex, 2-oxoglutarate dehydrogenase. The folding mechanism of BBL is under debate between the views of noncooperative downhill-type and classical two-state. Extensive replica exchange molecular dynamics simulations of BBL in explicit solvent have shown some...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2011.03.058

    authors: Fan J,Duan M,Li DW,Wu H,Yang H,Han L,Huo S

    更新日期:2011-05-18 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

    doi:10.1016/j.bpj.2009.06.011

    authors: Gliko O,Saggau P,Brownell WE

    更新日期:2009-08-19 00:00:00

  • A hydrodynamic theory of bilayer membrane formation.

    abstract::A simple formula for the expansion rate U of the bilayer membrane perimeter was derived by the solution of the hydrodynamic and the Laplace equations at small rates and small contact angles: U = theta delta/6 micron ln (h/e 3/2 h0), where theta is the equilibrium contact angle between the bilayer membrane and the Plat...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(81)84714-5

    authors: Dimitrov DS

    更新日期:1981-10-01 00:00:00