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At least 649 records · Page 36Linked to original sources
PubMed1979

Structural changes of ultraviolet-irradiated DNA derived from hydrodynamic measurements.

The paper reports on sedimentation and viscosity measurements performed on ultraviolet-irradiated DNA from T7 phage and calf thymus. From the hydrodynamic data the relative changes in the mean molecular weight, radius of gyration, and effective Kuhn statistical segment length were calculated. The results show that ultraviolet irradiation (254 nm) leads to a significant decrease of the effective statistical segment length of DNA which may be due to small local helix kinks (produced by the generation of photodimers) and a local increase of chain flexibility. Alterations in the overall DNA conformation may be observed even at low fluences where the mean molecular weight almost stays constant. The locally distorted helix regions possibly may serve as recognition sites in the first step of excision repair.

Animals↗
PubMed1978

Hydrodynamic studies on the self association of vertebrate skeletal muscle myosin.

Sedimentation velocity studies on myosin A solutions at high ionic strength combined with computer-simulation of the concentration dependence of the sedimentation coefficient for a rapidly reversible monomer-dimer equilibrium have confirmed that if such an equilibrium does exist it has an equilibrium constant of less than 10 ml/g. A new hydrodynamic treatment has been used to calculate the molecular weight of myosin from s0 and ks alone and has yielded a value of 470 000. Combination of viscosity and sedimentation velocity results has shown that the myosin molecule displays little swelling (Vs/v = 1.1 +/- 0.1). A new picture of the myosin molecule is presented in which a conformational change in the head region is suggested to account for the variation in published s 0 values.

Animals↗
PubMed1981

Hydrodynamic characterization of the heterodimer of spectrin.

The hydrodynamic properties of the spectrin dimer have been examined. The S20,w value of 9.3 S and the D20,w value of 1.75 x 10(-7) cm2 x s-1 yield a molecular weight of 470 000, in good agreement with the value from sedimentation equilibrium of 460 000. The frictional ratio of 2.3 and the intrinsic viscosity of 36 ml/g are not consistent with a compact, globular structure, but the value of the parameter Ks/[eta], 1.15, is not consistent with a rigid rod model. The most appropriate model, consistent also with data from other laboratories, is of a flexible, kinked rod-like molecule.

Diffusion↗
PubMed1981

Effect of chymosin action on the hydrodynamic diameter of casein micelles.

Quasi-elastic light scattering shows an initial decrease of about 5 nm in the hydrodynamic radius of casein micelles after adding chymosin, assuming the decrease to be equal for all micelles. This is consistent with the hypothesis that casein micelles have a hairy outer layer that is partly made up of the caseino-macropeptide part of kappa casein.

Caseins↗
PubMed1985

Hydrodynamic properties of a thromboxane A2/prostaglandin H2 antagonist binding site solubilized from human platelets.

The hydrodynamic properties of a binding site for the thromboxane A2/prostaglandin H2 receptor antagonist 9,11-dimethylmethano-11, 12-methano-16-(3-iodo-4-hydroxyphenyl)-13, 14-dihydro-13-aza-15 alpha beta-omega-tetranor-thromboxane A2 (I-PTA-OH) were determined in solubilized membrane proteins from human platelets using the detergent 3-[(3-cholamidopropyl)-dimethylammonio] 1-propane-sulfonate (CHAPS). Gel filtration revealed a Stokes radius of 5.25 +/- 0.37 nm (n=9). Molecular weight determined by gel filtration assuming a spherical protein was 180,000-220,000 Daltons. Sedimentation through sucrose or glycerol gradients revealed a sedimentation coefficient of 6.3 +/- 0.2 Svedberg units (n=5). The molecular weight calculated using the Stokes radius and sedimentation coefficient was 140,000 Daltons. The frictional ratio f/fo was 1.4, corresponding to an axial ratio of 7:1.

Binding Sites↗
PubMed1979

A computer model for hydrodynamic shearing of DNA.

A computer simulation model for the hydrodynamic shearing of DNA is presented. In this model the event space consists of a number of positions at which the DNA strand may break. The results of the model compared with the real world result are reported in this paper. Future endeavors to generalize the model would add significantly to the model's value.

Computers↗
PubMed1992

Active drag, useful mechanical power output and hydrodynamic force coefficient in different swimming strokes at maximal velocity.

By comparing the time of the same distance swum with and without an added resistance, under the assumption of an equal power output in both cases, the drag of 73 top swimmers was estimated. The active drag Fr(a.d.) at maximal swimming velocities varied considerably across strokes and individuals. In the females Fr(a.d.) ranged from 69.78 to 31.16 N in the front-crawl, from 83.04 to 37.78 N in dolphin, from 93.56 to 45.19 N in breaststroke, and from 65.51 to 37.79 N in back-stroke. In the males Fr(a.d.) ranged from 167.11 to 42.23 N in front-crawl, from 156.09 to 46.95 N in dolphin, from 176.87 to 55.61 N in breaststroke, and from 146.28 to 46.36 N in back-stroke. Also, the ratio of Fr(a.d.) to the passive drag Fr(a.d.) as determined for the analogical velocity in a tugging condition (in standard body position-front gliding) shows considerable individual variations. In the female swimmers variations in Fr(a.d.)/Fr(p.d.) ranged from 145.17 to 59.94% in front-crawl, from 192.39 to 85.57% in dolphin, from 298.03 to 124.50% in breaststroke, and from 162.87 to 85.61% in back-stroke. In the male swimmers variations in Fr(a.d.)/Fr(p.d.) ranged from 162.24 to 62.39% in front-crawl, from 191.70 to 70.38% in dolphin, from 295.57 to 102.83% in breaststroke, and from 198.82 to 74.48% in back-stroke. The main reason for such variations is found in the individual features of swimming technique and can be quantitatively estimated with the hydrodynamic force coefficient, which thus provides an adequate index of technique.

Adolescent↗
PubMed1993

Studies on new cross-axis coil planet centrifuge for performing counter-current chromatography. III. Speculations on the hydrodynamic mechanism in stationary phase retention.

Retention of the stationary phases of one organic-aqueous solvent system and three aqueous-aqueous polymer solvent systems was investigated on a cross-axis coil planet centrifuge. A graphic statistical treatment of all the data highlighted the prevailing effect of the inward-outward elution mode. A simplified model was proposed and studies on the paths and accelerations of cross-axis devices described in the previous paper provided explanations about the observed hydrodynamic behaviors.

Centrifugation↗
PubMed1996

Potential of on-line isotachophoresis-capillary zone electrophoresis with hydrodynamic counterflow in the analysis of various basic proteins and recombinant human interleukin-3.

Isotachophoresis-capillary zone electrophoresis with hydrodynamic counterflow was applied to the analysis of various basic model proteins and recombinant human interleukin-3. The use of coated capillaries and simple modifications of a commercial capillary zone electrophoresis (CZE) system allowed the detection of concentrations in the 10(-8) M region. The limit of detection was decreased by a factor of 100 compared to conventional CZE. The separation efficiency was unaffected by the preceding isotachophoretic step. Our method is applicable to the analysis of proteins with an isoelectric point higher than 7.0.

Buffers↗
PubMed1987

Structure and hydrodynamic properties of plectin molecules.

Plectin is a cytoskeletal, high molecular weight protein of widespread and abundant occurrence in cultured cells and tissues. To study its molecular structure, the protein was purified from rat glioma C6 cells and subjected to chemical and biophysical analyses. Plectin's polypeptide chains have an apparent molecular weight of 300,000, as shown by one-dimensional sodium dodecyl sulfate/polyacrylamide electrophoresis. Cross-linking of non-denatured plectin in solution with dimethyl suberimidate and electrophoretic analyses on sodium dodecyl sulfate/agarose gels revealed that the predominant soluble plectin species was a molecule of 1200 X 10(3) Mr consisting of four 300 X 10(3) Mr polypeptide chains. Hydrodynamic properties of plectin in solution were obtained by sedimentation velocity centrifugation and high-pressure liquid chromatography analysis yielding a sedimentation coefficient of 10 S and a Stokes radius of 27 nm. The high f/fmin ratio of 4.0 indicated a very elongated shape of plectin molecules and an axial ratio of about 50. Shadowing and negative staining electron microscopy of plectin molecules revealed multiple domains: a rigid rod of 184 nm in length and 2 nm in diameter, and two globular heads of 9 nm diameter at each end of the rod. Circular dichroism spectra suggested a composition of 30% alpha-helix, 9% beta-structure and 61% random coil or aperiodic structure. The rod-like shape, the alpha-helix content as well as the thermal transition within a midpoint of 45 degrees C and the transition enthalpy (168 kJ/mol) of secondary structure suggested a double-stranded, alpha-helical coiled coil rod domain. Based on the available data, we favor a model of native plectin as a dumb-bell-like association of four 300 X 10(3) Mr polypeptide chains. Electron microscopy and turbidity measurements showed that plectin molecules self-associate into various oligomeric states in solutions of nearly physiological ionic strength. These interactions apparently involved the globular end domains of the molecule. Given its rigidity and elongated shape, and its tendency towards self-association, plectin may well be an interlinking element of the cytoskeleton that may also form a network of its own.

Animals↗
PubMed1984

Hydrodynamic and biochemical correlates of the activation of the glucocorticoid-receptor complex.

Possible changes in the size and shape of the glucocorticoid-receptor complex (GRC) following activation remain poorly documented, due to the lability and possible activation of the receptor during the determination of these hydrodynamic parameters. In the present study molybdate was used to stabilize the GRC, thus preventing these uncontrolled transformations. Cytosol prepared from mouse whole brains was incubated for 18 h at 0-2 degrees C with [3H]triamcinolone acetonide (+/- molybdate). Activation was then initiated by incubation at 22 degrees C for variable times and quenched at 0 degree C by adding molybdate. The Stokes radius and sedimentation coefficient of the GRC declined from 77 A and 9.2 S before activation to 58 A and 3.8 S after activation. These measurements remained consistent after recycling GRC between sedimentation and gel filtration procedures and correspond to a 3-fold reduction in the relative molecular mass. The loss and formation of the 297 and 92 kDa species, respectively, after different durations of activation correlated nearly perfectly with increased binding of GRC to DNA-cellulose (DNA-C). The observed size change also correlated well with decreased adsorption to DEAE-cellulose filters (DE-81) and increased adsorption to glass fiber filters (GF/C). The increased adsorption to GF/C may reflect an increase in hydrophobicity which, with extended durations of activation, leads to increased aggregation and reduced binding to DNA-C, but not to a change in adsorption to DE-81. We propose that during activation the 297 kDa form of the GRC splits to form a 92 kDa species that displays an increased affinity for DNA.

Animals↗
PubMed1984

Ex vivo human platelet aggregation induced by decompression during reduced barometric pressure, hydrostatic, and hydrodynamic (Bernoulli) effect.

Decompression of human platelet-rich plasma (PRP) in siliconized glass or plastic to 380 mm Hg for 3 hours at 38 degrees C produced platelet aggregation independent of pO2. Aggregation also took place when PRP was compressed to 8,000 PSI and then decompressed slowly to one atmosphere (14.7 PSI) without gas bubble formation. Platelets also aggregated when plasma was decompressed hydrodynamically (Bernoulli effect) at room temperature. It was also found that the drugs piracetam (2-oxypyrolidine acetamide) and pentoxifylline (1-(5-oxohexyl)-theobromine) at 0.5 and 1.0 mM prevent thrombocyte aggregation. Implications for mountain sickness are discussed.

Atmospheric Pressure↗
PubMed1989

Structural differences between histone H1 molecules from sea urchin (Strongylocentrotus intermedius) sperm and calf thymus: hydrodynamic and c.d. studies.

Comparative sedimentation, diffusion and circular dichroism (c.d.) measurements have been performed on two histones H1 from sperm of the sea urchin Strongylocentrotus intermedius (H1S) and from calf thymus (H1T), at a high salt concentration of M NaCl. Both the Stokes radius and the frictional ratio derived from the hydrodynamic parameters were found to be somewhat smaller for H1S than the corresponding values for H1T. In view of the considerably higher molar mass of H1S compared with that of H1T, this result indicates that H+S in 2 M NaCl has a more compact conformation than H1T, probably due to a higher degree of secondary structure in the flanking domains of H1S. The c.d. measurements likewise show that H1S has a higher content of ordered structures than H1T. Model considerations indicate that the C-terminal tail of H1S is the main candidate for accommodation of these additional secondary structure regions.

Animals↗
PubMed1991

Analysis of hydrodynamic data for denatured globular proteins in terms of the wormlike cylinder model.

Hydrodynamic data, i.e. intrinsic viscosity and sedimentation coefficient, for denatured globular proteins in 6 M guanidinium chloride have been re-analysed in terms of the Yamakawa-Fujii theory of the wormlike cylinder model. Molecular parameters thus obtained ((mean value of R2 0/M) infinity, the Kuhn statistical segment length and the molecular weight per unit contour length) are in better agreement with the values obtained theoretically or by other methods than those evaluated on the basis of the model of non-draining random coil.

Mathematics↗
PubMed1993

Hydrodynamic characterization of chitosans varying in degree of acetylation.

The molecular weights and gross aqueous solution conformation of two chitosans of different degrees of acetylation, 'Sea Cure +210' (11% acetylated) and KN50 (58% acetylated) were characterized by viscometry, analytical ultracentrifugation and dynamic light scattering. The hydrodynamic parameters obtained were used to determine both the molecular weights and the gross solution conformation of the two chitosans. Using the Wales-Van Holde ratio of sedimentation coefficient concentration regression coefficient (ks) to intrinsic viscosity [eta], the Sea Cure +210 chitosan, which is much less acetylated than the KN50, is highly asymmetric in conformation. This, in conjunction with the charge on the molecule, would suggest a rod-like conformation in solution. The two largest KN50 chitosans have widely differing values for the Wales-Van Holde ratio, suggesting different solution conformation. However, when the series is examined as a whole using the Mark-Houwink-Kuhn-Sakurada relationships relating molecular weight to both intrinsic viscosity and translational diffusion coefficient, then a more spheroidal structure, approximating to a random coil, is predicted.

Acetylation↗
PubMed1996

The interaction of sodium dodecyl sulfate and urea with cat-fish collagen solutions in acetate buffer: hydrodynamic and thermodynamic studies.

Cat-fish collagen was extracted and characterized. Shrinkage temperature of cat-fish collagen is 54.5 degrees C. SDS-PAGE pattern indicated that the cat-fish collagen is Type I in nature. The ratio of proline and hydroxyproline is 1:2 and it suggests cat-fish collagen is vertebrate. The molecular weight of cat-fish collagen was determined by using molecular sieve chromatography and it was found to be 3 20,000 Da. The mutual interaction of cat-fish collagen with SDS and urea was studied at various temperatures. The results suggest that the aggregation of collagen is facilitated by the presence of SDS, whereas hindered by urea. The various thermodynamic parameters were estimated from viscosity measurements and the transfer of collagen into SDS micelles, urea and the reverse phenomenon was analysed. These transfer properties are temperature-dependent. Our thermodynamic results are also able to predict the exact denaturation temperature as well as the structural order of water in the collagen in various environments. The hydrated volumes, Vh of collagen in buffer, SDS, and urea environments using Simha-Einstein equation and intrinsic viscosity were also calculated. The low intrinsic viscosity [eta] and high Vh value of collagen in an SDS environment compared to buffer and other environments suggested a more workable system in cosmetic and dermatological preparations. The one and two-hydrogen-bonded models of this collagen in various environments have been analysed. The calculated thermodynamic parameters varied with the concentration of collagen as well as concentration of additives. The change of thermodyanamic parameters from coiled-coil to random-coil conformation upon denaturation of collagen were calculated from the amount of proline and hydroxyproline residues and compared with viscometric results. Denaturation enthalpy of the catfish collagen in buffer, SDS and urea environments has also been determined by differential scanning calorimetric (DSC) measurements, and the results are in good agreement with the viscosity-derived values. The assymmetry and molecular geometry of this collagen in buffer, SDS and urea environments are also computed. Overall, our hydrodynamic and thermodynamic results suggest that the stability of the collagen in the additive environments is in the following order: SDS > buffer > urea.

Acetates↗
PubMed1987

Hydrodynamic characterization of the size and shape of atropinesterase from Pseudomonas putida.

Atropinesterase from Pseudomonas putida has been investigated by means of different ultracentrifugation methods under native and denaturing conditions. The following quantities were determined: sedimentation coefficient, translational diffusion and friction coefficient, partial specific volume and molecular weight. From these data the size, shape and hydration of the enzyme molecule in solution were estimated. The results suggest that atropinesterase is a globular protein which consists of a single polypeptide chain with a molecular weight of about 30,000. In solution under non-denaturing conditions, it occurs mainly as a dimer which hydrodynamically behaves as a rigid impenetrable particle. Calculations based on the spheroid model indicate that this particle resembles a hydrated sphere with a diameter of 6.1 +/- 0.2 nm and a hydration of 0.4 +/- 0.1 g of H2O/g of protein rather than a significantly less hydrated ellipsoid of revolution. Under denaturing conditions dissociation into monomers takes place. The effects of sodium dodecyl sulphate (SDS) on size and shape suggest that dimerization results from side-by-side association of two elongated monomers rather than from end-to-end association. Approximately 57 molecules of SDS are bound per dimer before dissociation occurs concomitant with the additional binding of about 19 molecules of detergent.

Carboxylic Ester Hydrolases↗
PubMed1990

The protective effect of serum against hydrodynamic damage of hybridoma cells in agitated and surface-aerated bioreactors.

The effect of serum on the growth rate and metabolism of CRL-8018 hybridoma cells in an agitated, surface-aerated bioreactor was examined. In the employed well-controlled bioreactors at high agitation rates, hybridoma cells in medium containing 1% fetal bovine serum rapidly die in the presence of a vortex with accompanying gas-bubble entrainment, whereas non-agitated control cultures grow normally in medium containing 1% serum. Serum levels greater than 5% counteract the detrimental hydrodynamic effects due to agitation and bubble entrainment. The protective effect is present after short-term (less than 1 h) exposure to 10% serum concentration, suggesting a protection mechanism which is, at least in part, of a physical nature. The apparent cell yields on glucose, lactate, and glutamine decreased with decreasing growth rate due to low serum concentrations. The results are incorporated into a simple model in which the apparent growth rate is the sum of an invariable growth rate and a changing death rate.

Animals↗