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At least 487 records · Page 27Linked to original sources
PubMed2003

Clearance rates of biotinylated ferritins in canine: a possible physiological role of canine serum ferritin as an iron transporter.

To elucidate the physiological role of canine serum ferritin, we measured clearance rates of biotinylated ferritins in beagle. Biotinylated canine tissue ferritins were cleared rapidly from circulation. The clearance time (T 1/2) of liver ferritin (H/L subunit ratio = 0.43) was 6.8 to 11.8 min, and that of heart ferritin (H/L = 3.69) was 9.3 to 25.0 min. T 1/2 of biotinylated canine liver ferritin was independentof iron content, whereas canine heart apoferritin (T 1/2 = 31.2 and 32.7 min) was more slowly removed from circulation than the holoferritin. On the other hand, biotinylated recombinant bovine H-chain ferritin homopolymer show a much slower rate of removal (T 1/2 = 153.8 and 155.0 min) compared with the L-chain ferritin homopolymer (T 1/2 = 26.4 and 31.3 min). The rapid clearance of canine tissue ferritin suggests that serum ferritin is an iron transporter in canines.

Animals↗
PubMed2003

Fracture toughness of tooth acrylics.

The hardness, fracture toughness, toughness, flexural strength and Young's moduli of three acrylic tooth polymers were investigated. The first polymer was based on a conventional homopolymer poly(methylmethacrylate). The second was based on cross-linked poly(methylmethacrylate) with an uncross-linked poly(methylmethacrylate) coating. The third material was based on an interpenetrating polymer network (IPN) of a cross-linked and uncross-linked poly(methylmethacrylate). All three polymers had similar hardness values. The cross-linked and IPN polymers had higher fracture toughness (K(IC)) and toughness (G(IC)) values than the conventional homopolymer poly(methylmethacrylate) polymer and lower flexural strengths (sigma(f)). The toughness of the cross-linked and IPN polymers was higher due to crack deflection around the polymer bead structure and the polymer beads acting as crack pinning sites.

Journal Article↗
PubMed2002

Biosynthesis of novel thermoplastic polythioesters by engineered Escherichia coli.

The development of non-petrochemical sources for the plastics industry continues to progress as large multinationals focus on renewable resources to replace fossil carbon. Many bacteria are known to accumulate polyoxoesters as water-insoluble granules in the cytoplasm. The thermoplastic and/or elastomeric behaviour of these biodegradable polymers holds promise for the development of various technological applications. Here, we report the synthesis and characterization of microbial polythioesters (PTEs), a novel class of biopolymers of general technological relevance. Biosynthesis of PTE homopolymers was achieved using a recombinant strain of Escherichia coli that expressed a non-natural pathway consisting of a butyrate kinase, a phosphotransbutyrylase, and a PHA synthase. Different homopolymers were produced, consisting of either 3-mercaptopropionate, 3-mercaptobutyrate, or 3-mercaptovalerate repeating units, if the respective mercaptoalkanoic acids were provided as precursor substrates to the fermentative process. The PTEs contributed up to 30% (w/w) of the cellular dry weight and were identified as hydrophobic inclusions in the cytoplasm. The chemical and stereochemical homogeneity of the purified PTEs were identified by different methods, and the estimated physical properties were compared to the oxypolyester equivalents, revealing low crystalline order and, for the poly(3-mercaptopropionate) improved thermal stability. The ability to produce PTEs through a biosynthetic route opens up new avenues in the field of biomaterials.

Biopolymers↗
PubMed2006

Raman spectroscopic study of the structure of water in aqueous solutions of amphoteric polymers.

Methacrylic acid (MA) and [3-(methacryloylamino)propyl]trimethylammonium chloride (MAPTAC) were polymerized to give amphoteric copolymers with various compositions. The structure and H-bonding of water in an aqueous solution of the copolymer were analyzed using the contours of the O-H stretching in the polarized Raman spectra. For comparison, the H-bonded network structure of aqueous solutions of homopolymers (polyMA and polyMAPTAC) was also examined. From the relative intensity of the collective band (C value) corresponding to a long range coupling of the O-H stretching in the aqueous polymer solutions, the number of H-bonds disrupted due to the presence of one monomer residue of the polymers (Ncorr) was determined. The Ncorr value for polyMA was largely positive, and with an increase in the content of the MAPTAC residue, the Ncorr value became smaller, and after passing a minimum (which was still slightly positive) at a roughly equivalent molar ratio, the Ncorr value increased again. This is in significant contrast with the larger positive Ncorr values for the homopolymers (both polyMA and polyMAPTAC), and other ordinary polyelectrolytes such as sodium polyethylenesulfonate, poly-L-lysine hydrobromide and sodium polyacrylate. Furthermore, the Ncorr value for the copolymer (MA ratio MAPTAC = 56:44) became much smaller by the neutralization of MA residues in the copolymer with sodium hydroxide, and comparable to those for neutral polymers such as poly(ethylene glycol) and poly(N-vinylpyrrolidone) and zwitterionic polymers such as poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC) and poly[3-sulfo-N,N-dimethyl-N-(3'-methacryloylaminopropyl)propanaminium inner salt]. The present results clearly indicate that the amphoteric polymers with comparative contents of cationic and anionic groups do not significantly disturb the H-bonded network structure of water, probably due to the counteraction of the electrostatic hydration effect by the proximity between the anionic and cationic side groups.

Journal Article↗
PubMed1968

Chain extension of ribonucleic acid by enzymes from rat liver cytoplasm.

1. The 105000g supernatant fraction of rat liver catalyses the incorporation of ribonucleotides from ribonucleoside triphosphates into polyribonucleotide material. The reaction requires Mg(2+) ions and is enhanced by the addition of an ATP-generating system and RNA, ATP, UTP and CTP but not GTP are utilized in this reaction. In the case of UTP, the product is predominantly a homopolymer containing 2-3 uridine residues, and there is evidence that these may be added to the 3'-hydroxyl ends of RNA or oligoribonucleotide primers. 2. The microsome fraction of rat liver incorporates ribonucleotides from ATP, GTP, CTP and UTP into polyribonucleotide material. This reaction requires Mg(2+) ions and is enhanced slightly by the addition of an ATP-generating system, and by RNA but not DNA. Supplementation of the reaction mixture with the three complementary ribonucleoside 5'-triphosphates greatly increases the utilization of a single labelled ribonucleoside 5'-triphosphate. The optimum pH is in the range 7.0-8.5, and the reaction is strongly inhibited by inorganic pyrophosphate and to a much smaller degree by inorganic orthophosphate. It is not inhibited by actinomycin D or by deoxyribonuclease. In experiments with [(32)P]UTP in the absence of ATP, GTP and CTP, 80-90% of (32)P was recovered in UMP-2' or -3' after alkaline hydrolysis of the reaction product. When the reaction mixture was supplemented with ATP, GTP and CTP, however, about 40% of the (32)P was recovered in nucleotides other than UMP-2' or -3'. Although the reactions seem to lead predominantly to the synthesis of homopolymers, the possibility of some formation of some heteropolymer is not completely excluded.

Adenosine Triphosphate↗
PubMed1972

Fractionation of nuclei from brain by zonal centrifugation and a study of the ribonucleic acid polymerase activity in the various classes of nuclei.

1. The nuclei of the cells of the whole rat brain have been fractionated in a B-XIV zonal rotor with a discontinuous gradient of sucrose. Five fractions were obtained. Zone (I) contained neuronal nuclei (70%) and astrocytic nuclei (23%). Zone (II) contained astrocytic nuclei (81%) and neuronal nuclei (15%). Zone (III) contained astrocytic nuclei (84%) and oligodendrocytic nuclei (15%). Zone (IV) contained oligodendrocytic nuclei (92%) and zone (V) contained only oligodendrocytic nuclei. 2. The content of DNA, RNA and protein per nucleus was determined for each zone. Although the amount of DNA per nucleus is constant (7pg) the RNA varies from 4.5 to 2.5pg/nucleus and the protein from 38 to 17.6pg/nucleus. The neuronal nuclei have the greatest amounts of protein. The oligodendrocytic nuclei have the least content of RNA and protein. 3. The effects of pH, ionic strength, and Mg(2+) and Mn(2+) concentration on the activity of the nuclear system for synthesis in vitro of RNA have been investigated for unfractionated nuclei. From these studies a standard set of conditions for the assay of nuclear RNA polymerase has been established. 4. The activity of the RNA polymerase in each of the zonal fractions has been determined in the presence and in the absence of alpha-amanitin. Zone (II) is the most active, followed by zone (I). The nuclei of zones (IV) and (V) have comparable activity, which is 40% of that of zone (II). 5. The extent of incorporation of each of the four labelled nucleoside triphosphates by the nuclei from each zone has been measured. These values have been used to calculate the base composition of the RNA synthesized in vitro in each class of nucleus. 6. The effect of changes in the condition of assay of RNA polymerase in the different classes of nuclei has been investigated. Significant differences in the response to concentrations of metal ions and ammonium sulphate have been observed. 7. Homopolymer formation in each zone of brain nuclei has been determined. The extent of formation of the four homopolymers roughly parallels the RNA polymerase activity.

Animals↗
PubMed1981

The influence of pressure on the self-assembly of the thick filament from the myosin of vertebrate skeletal muscle.

The thick-filament-monomeric-myosin equilibrium was prepared from pure myosin at pH 8.1. The application of hydrostatic pressure to the self-assembly equilibrium resulted in a biphasic dissociation curve in which a linear decrease in turbidity (a measure of weight added to or lost from the filament) was followed by a transition to a second pressure-insensitive phase. The first phase represents the effect of hydrostatic pressure on the growth or propagation phase of filament assembly. Here is was shown that hydrostatic pressure served to shorten the filaments in concert towards the bare zone whilst maintaining the narrow length distribution seen at atmospheric pressure; the filament concentration remained constant during the experiment. A more precise definition of the delta-v for the assembly of monomer into filament was obtained than had hitherto been possible. The positioning of the bare zone at the centre of the filament seems to be one of the more obvious functions of the length-regulation mechanism. It also appears that all the basic structural elements of the native thick filament are potentially present in the pH 8.1 homopolymer; its length can be increased by physiological concentrations of MgCl2 and decreased by pressure. The monodisperse native filament could then be formed by a fine tuning of the basic length-regulation mechanism of the homopolymer by the co-polymerization of the other thick-filament proteins.

Animals↗
PubMed1992

Evidence of H- and L-chains have co-operative roles in the iron-uptake mechanism of human ferritin.

The ability to incorporate iron in vitro was studied in homopolymers of human ferritin L-chain, human ferritin H-chain and its variants and in homopolymer mixtures. The H-chain variants carried amino acid substitutions in the ferroxidase centre and/or in carboxy residues on the cavity surface. Iron incorporation was examined by gel electrophoresis of the reaction products by staining for iron and protein. It was found that inactivation of the ferroxidase centre combined with the substitution of four carboxy groups on the cavity abolished the ability of H-chain ferritin to incorporate iron. Competition experiments with limited amounts of iron showed that, at neutral pH, L-chain ferritin is more efficient in forming iron cores than the H-chain variants altered at the ferroxidase activity or in the cavity. Competition experiments at pH 5.5 demonstrated that L-chain apoferritin is able to incorporate iron only when in the presence of H-chain variants with ferroxidase activity. The results indicate that L-chain apoferritin has a higher capacity than the H-chain apoferritin to induce iron-core nucleation, whereas H-chain ferritin is superior in promoting Fe(II) oxidation. The finding of cooperative roles of the H- and L-chains in ferritin iron uptake provides a clue to understanding the biological function of isoferritins.

Apoferritins↗
PubMed1998

Transient overexpression of human H- and L-ferritin chains in COS cells.

The understanding of the in vitro mechanisms of ferritin iron incorporation has greatly increased in recent years with the studies of recombinant and mutant ferritins. However, little is known about how this protein functions in vivo, mainly because of the lack of cellular models in which ferritin expression can be modulated independently from iron. To this aim, primate fibroblastoid COS-7 cells were transiently transfected with cDNAs for human ferritin H- and L-chains under simian virus 40 promoter and analysed within 66 h. Ferritin accumulation reached levels 300-500-fold higher than background, with about 40% of the cells being transfected. Thus ferritin concentration in individual cells was increased up to 1000-fold over controls with no evident signs of toxicity. The exogenous ferritin subunits were correctly assembled into homopolymers, but did not affect either the size or the subunit composition of the endogenous heteropolymeric fraction of ferritin, which remained essentially unchanged in the transfected and non-transfected cells. After 18 h of incubation with [59Fe]ferric-nitrilotriacetate, cellular iron incorporation was similar in the transfected and non-transfected cells and most of the protein-bound radioactivity was associated with ferritin heteropolymers, while H- and L-homopolymers remained iron-free. Cell co-transfection with cDNAs for H- and L-chains produced ferritin heteropolymers that also did not increase cellular iron incorporation. It is concluded that transient transfection of COS cells induces a high level of expression of ferritin subunits that do not co-assemble with the endogenous ferritins and have no evident activity in iron incorporation/metabolism.

Animals↗
PubMed1998

Purification and characterization of a 100 kDa DNA polymerase from cauliflower inflorescence.

A DNA polymerase from cauliflower (Brassica oleracea var. botrytis) inflorescence has been purified to near homogeneity through five successive column chromatographies, and temporally designated cauliflower polymerase 1. Cauliflower polymerase 1 is a monopolypeptide with a molecular mass of 100 kDa. The enzyme efficiently uses synthetic DNA homopolymers and moderately activated DNA and a synthetic RNA homopolymer as template-primers. The enzyme is strongly sensitive to dideoxythymidine triphosphate and N-ethylmaleimide, but it is insensitive to aphidicolin. It was stimulated with 250 mM KCl. Its mode of DNA synthesis is high-processive with or without proliferating-cell nuclear antigen. A 3'-->5' exonuclease activity is associated with cauliflower polymerase 1. The enzyme is clearly different from cauliflower mitochondrial polymerase and does not resemble the four different types of wheat DNA polymerase, designated wheat DNA polymerases A, B, CI and CII. In the present paper the role of the enzyme in plant DNA synthesis is discussed.

Brassica↗
PubMed2001

Biosynthesis of PHB tercopolymer by Bacillus cereus UW85.

AIMS: The study was attempted to determine the ability of a Gram-positive Bacillus cereus UW85 strain to biosynthesize poly (3-hydroxybutyrate) copolymers when epsilon-caprolactone, or epsilon-caprolactone and glucose, were used as carbon sources. METHODS AND RESULTS: Bacillus cereus was grown for 24 h under nitrogen-limited conditions in a mineral salts medium. Growth was monitored by measurement of turbidity. Glucose level was determined by the colorimetric anthrone METHOD: The epsilon-caprolactone concentration was determined by gas chromatography. The bacterial biopolymers were extracted with chloroform in a Soxhlet extractor and then characterized by nuclear magnetic resonance and gel permeation chromatography. When epsilon-caprolactone was used as a carbon substrate, the bacterial strain produced tercopolymer with 3-hydroxybutyrate, 3-hydroxyvalerate and 6-hydroxyhexanoate units. However, when caprolactone and glucose were supplied together, only homopolymer of poly (3-hydroxybutyrate) was produced. CONCLUSION: All tercopolymers isolated from B. cereus UW85 cells were obtained with yields up to 9% (w/w) and low number-average molecular weight compared with the homopolymer PHB. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacillus cereus UW85 produced tercopolymer with a low molecular weight from one substrate (epsilon-caprolactone) used as a carbon source. The results are significant for the potential future application of Bacillus biopolymers to bioplastics production.

Bacillus cereus↗
PubMed2005

Effects of triplet exciton confinement induced by reduced conjugation length in polyspirobifluorene copolymers.

Using gated optical spectroscopy at low temperatures, a polyspirobifluorene has been compared with an alternating carbazole-spirobifluorene copolymer in which the backbone conjugation is interrupted due to meta coupling of the carbazole moieties. In the copolymer both singlet and triplet energy levels are blueshifted by 130 meV with respect to the homopolymer, resulting in an unaltered singlet-to-triplet splitting. Though the barrier for triplet exciton migration increases from 4.4 to 6.0 meV for the copolymer compared to the homopolymer, it still remained low enough to ensure efficient triplet diffusion at ambient temperature.

Journal Article↗
PubMed2006

Equilibrium and dynamics of Langmuir monolayers when the interface is a selective solvent: Polystyrene-b-poly(t-butyl acrylate) block copolymers.

The surface pressure of monolayers of insoluble diblock copolymers has been measured. One of the blocks is made of poly(t-butyl acrylate) (PtBA), and the other one by polystyrene (PS). The interface is a good solvent for PtBA, while it is a poor solvent for PS. For the sake of comparison, monolayers of a PtBA homopolymer (good solvent conditions) and of poly(4-hydroxy styrene) (P4HS) (poor solvent conditions) have been also measured. It has been found that the relative length of the blocks plays an important role on the shape of the surface pressure Pi versus surface concentration Gamma curves and also on the shape of the equilibrium compressibility versus Gamma curves. However, it does not affect the maximum value of Pi reached at high Gamma's. Surprisingly, the ellipsometric thickness of the copolymer monolayers is almost independent of the relative length of the blocks. The dynamics of the monolayers has been studied by step compression and by surface-light scattering techniques. When M(w,PtBA) >> M(w,PS) single exponential relaxations are observed. However, stretched exponentials are obtained for M(w,PS) > or = M(w,PtBA). The relaxation times decrease with increasing Gamma for all the copolymers studied. This is the behavior usually found for poor solvent conditions (P4HS) and opposite to that found for homopolymers under good solvent conditions [PtBA, poly(vinylacetate)]. This means that the solvent quality of the interface does not determine the pressure dependence of tau. The elasticity modulus of the monolayers in the kilohertz range takes values that are similar to those of the high-frequency limit of the relaxation experiments. This means that the relaxation processes have characteristic frequencies below 1 Hz.

Journal Article↗
PubMed2006

Adsorption of diblock copolymers on stripe-patterned surfaces.

We present the results of extensive Monte Carlo simulations of diblock copolymers adsorbed on stripe-patterned surfaces of various widths. We have found that the width of the stripe pattern is an important parameter which dictates favorable recognition on the surface. For certain stripe widths, the adsorption of diblock copolymers to striped surfaces exhibits two transitions. The process involves recognition of the surface pattern by the diblock copolymer which follows a two step process in which the first block getting adsorbed to the appropriate pattern on the surface, without any recognition of the surface pattern, followed by the adsorption of the second block, where a reorganization process happens. For small widths and also for higher widths, the chain behaves just like a homopolymer where the twofold adsorbing process changes to the typical homopolymer adsorption. We have also found that there exists an optimal width of the stripes, independent of the chain length, where the recognition on the surface pattern is most favored. The characteristic temperature of the adsorption of the second block with weaker interactions is found to be independent of the chain length at this optimal width, proving that only local rearrangements take place after the first step. Some of our results describing the thermodynamics compare very well with the recent semianalytical approach of Kriksin et al. [J. Chem. Phys. 122, 114703 (2005)] on multiblock copolymers on heterogeneous surfaces. We also present some interesting conformational properties of the copolymer chain near the stripe-patterned surface.

Journal Article↗
PubMed2006

Density functional theory for the recognition of polymer at nanopatterned surface.

The recognition of homopolymer at nanopatterned surface has been investigated by density functional theory (DFT). Chain conformation and pattern transfer parameter predicted from the DFT are in good agreement with Monte Carlo simulation results. The theory describes satisfactorily the transition from depletion at low packing fractions to adsorption and double-layer adsorption at high packing fractions and also accounts for the crucial effect of the segment-wall interaction. It is found that homopolymer is better recognized at a low bulk density and a stronger interaction with the surface. The polymer can not only recognize the surface but also invert the surface at high bulk densities. The chain in the solution-wall interface exhibits a typical "brush" conformation with a length approximated by half the length of polymer chain.

Journal Article↗
PubMed2000

Reelin molecules assemble together to form a large protein complex, which is inhibited by the function-blocking CR-50 antibody.

Reelin is a key mediator of ordered neuronal alignment in the brain. Here, we demonstrate that Reelin molecules assemble with each other to form a huge protein complex both in vitro and in vivo. The Reelin-Reelin interaction clearly is inhibited by the function-blocking anti-Reelin antibody, CR-50, at a concentration known to inhibit Reelin function. This assembly is mediated by electrostatic interaction of the CR-50 epitope region. Recombinant CR-50 epitope fragments spontaneously constitute a soluble, string-like homopolymer with a regularly repeated structure composed of more than 40 monomers. Mutated Reelin, which lacks the CR-50 epitope region, cannot form a homopolymer and fails to induce efficient tyrosine phosphorylation of Disabled 1 (Dab1), which should occur to transduce the Reelin signal. These data suggest that Reelin exerts its biological function by composing a large protein assembly driven by the CR-50 epitope region, proposing a novel model of the Reelin signaling in neurodevelopment.

Animals↗
PubMed1969

Role of base-pairing in the control of an enzyme reaction.

In an attempt to ascertain whether specific base-pairing could have a regulating function in an enzyme system in which nucleotides serve as the substrate, we studied the effects of the addition of the various homopolymers of the ribonucleotide series on the action of adenylate kinase on ADP or CDP, and found that the results support this possibility. Poly U strongly inhibits the enzyme action on ADP, and poly G that on CDP, both to the extent of about 70 per cent. Lesser effects are shown when poly G is used with ADP and poly U with CDP. The other homopolymers tested, poly A and poly C, are ineffective.

Adenine Nucleotides↗
PubMed1970

Oxidized RNase as a protein model having no contribution to the hydrogen exchange rate from conformational restrictions.

As oxidized RNase is a model for the random conformation state of RNase, the hydrogen exchange kinetics of oxidized RNase approximate the intrinsic conformation-independent chemical exchange rate of the native protein. The energy of activation, the pH(min), and the k(min) of oxidized RNase exchange rates are similar to those reported for amino acid homopolymers. However, unlike the exchange from homopolymers, the exchange from oxidized RNase is characterized by a distribution of first-order rates. This distribution is important to the analysis of exchange from native proteins in terms of classes of sites which share common structural properties.

Animals↗