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  • 1
    ISSN: 1432-1017
    Keywords: Trimethylamine dehydrogenase ; Analytical ultracentrifugation ; Hydrodynamics ; Homodimers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Physics
    Notes: Abstract Earlier studies using x-ray crystallography have shown that trimethylamine dehydrogenase (TMADH) from methylotropic bacteria exists as homodimers in the crystalline state. In this present hydrodynamic study we show that this is true also in dilute solution conditions and investigate the degree of swelling or relaxation of the protein in solution. Analytical ultracentrifugation was used to determine the molar mass and to investigate whether the homodimeric nature of this molecule in crystal form — as visualized by x-ray crystallography — is reproduced in dilute solution at temperatures between 4 and 40°C. The globular solution structure determined at 4 and 40°C is in good agreement with crystallographic data although trimethylamine dehydrogenase was found to be either more asymmetric in solution — or highly hydrated —, a phenomenon found to increase with temperature. In agreement with the crystallographic structure, the enzyme sediments as a homodimer with a molar mass of (163,000±5,000) g/mol. The concentration dependence of the sedimentation coefficient in the range of 0–1 mg/ml, indicates that no association or dissociation occurs. These findings are additionally supported by sedimentation equilibrium data in the concentration range of 0 to 1.8 mg/ml. Finally, from the sedimentation coefficient distribution at various temperatures, it was concluded that the enzyme is conformationally flexible and assumes an even more expanded structure at higher temperatures which is in good agreement with the hydrodynamic calculations performed.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1022-1352
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Physics
    Notes: Small block copolymers consisting of a hydrophilic poly(ethylene glycol) block and a second, also hydrophilic, moiety which strongly interacts with alkaline earth ions were synthesized by diverse reaction sequences based on poly(ethylene glycol) monomethyl ethers (MW = 2000 and 5000 g/mol, respectively). These starting blocks were transferred to the poly(ethylene glycol) monomethyl monoglycidyl ether or to poly(ethylene glycols) with one terminal acid chloride group. Both intermediates were subsequently reacted with poly(ethyleneimine) (MW = 700 g/mol) and bromoacetic acid to yield poly(ethylene glycol)-block-poly[(N-carboxymethyl)ethyleneimine] (PEG-b-PEIPA, average MW = 3800 resp. 6800 g/mol) as a polymeric analog of EDTA. The terminal epoxy group is also convenient for the connection of simple pèptide sequences. For the desired purpose of ion binding, poly(aspartic acid) (PAsp) was applied, resulting in the block copolymer PEO-b-PAsp. A simple testing procedure concerning the inhibition of calcium carbonate precipitation was applied for the prepared structures. A comparison with commercial builders for water treatment such as poly[(acrylic acid)-co-(maleic anhydride)] or poly(aspartic acid) stresses the superb calcium carbonate crystallization inhibition efficiency (up to the 20 fold) of the double hydrophilic block copolymer stabilizers.
    Additional Material: 2 Tab.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 0006-3525
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: A laboratory-made sample of the polysaccharide xylinan (acetan) has been further characterized with respect to (i) purity, (ii) molar mass and polydispersity, and (iii) gross conformation by a combination of hydrodynamic measurements (sedimentation velocity and equilibrium analytical ultracentrifugation, viscometry, and dynamic light scattering) in aqueous NaCl (I = 0.10 mol·L-1). Sedimentation velocity diagrams recorded using Schlieren optics revealed highly pure material sedimenting as a single boundary [so20.w = 9.5 ± 0.7) S; ks = (273 ± 112) mL/g]. The hypersharp nature of these boundaries is symptomatic of a polydisperse and highly nonideal (in the thermodynamic sense) system. Low speed sedimentation equilibrium in the analytical ultracentrifuge using Rayleigh interference optics and two different types of extrapolation procedure (involving point and whole-cell molar masses) gave a weight average molar mass Mw of (2.5 ± 0.5) × 10-6 g·mol-1 and also a second virial coefficient, B = (2.8 ± 0.7) × 10-4 mL·mol·g-2, both values in good agreement with those from light scattering-based procedures (Part II of this series). A dynamic Zimm plot from dynamic light scattering measurements gave a z-average translational diffusion coefficient Do20.w = (3.02 ± 0.05) × 10-8 cm2·s-1 and the concentration-dependence parameter kD = (370 ± 15) mL/g. Combination of so20.w with Do20.w via the Svedberg equation gave another estimate for Mw of ≅ 2.4 × 106 g/mol, again in good agreement. Both the Wales-van Holde ratio (ks/[η]) ≅ 0.4 (with [η] = (760 ± 77) mL/g) and the ρ-parameter (ratio of the radius of gyration from static light scattering to the hydrodynamic radius from dynamic light scattering) as ρ 〉 2.0 all indicate an extended conformation for the macromolecules in solution. These findings, plus Rinde-type simulations of the sedimentation equilibrium data are all consistent with the interpretation in terms of a unimodal wormlike coil model performed earlier. © 1996 John Wiley & Sons, Inc.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Chemie in unserer Zeit 33 (1999), S. 200-205 
    ISSN: 0009-2851
    Keywords: Chemistry ; Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Amphiphilic block copolymers are versatile structure-directing agents for the synthesis of organic nanostructured materials. Chemically well characterized and easily accessible, they can be utilized for a rich variety of applications. While the prefabrication of an ABC “nanoreactor” or a mesoscopic casting mold occur via spontaneous self-assembly (exo-and endo-templating), during biomimetic mineralization the structure of the resulting inorganic-ABC hybrid material is induced upon combining thc components. The underlying methodologies are illustrated using recently published, representative examples.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Macromolecular Chemistry and Physics 196 (1995), S. 3469-3485 
    ISSN: 1022-1352
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Physics
    Notes: If gels are investigated in an analytical ultracentrifuge by means of sedimentation equilibrium experiments, so called soluble parts which are originally not incorporated into the network influence the swelling pressure equilibria of the gels. For gelatin/water, it can be shown that an irreversible process takes place which is dependent on the rotational speed of the experiment. Experimental evidence is given that soluble parts which are partly associated and aggregated to the polymer network can act as a branching and/or crosslinking agent. A model for this process is given. In such a case an application of the results of the swelling theory, e.g. that of Flory and Huggins assuming a constant number of network junctions, cannot be applied anymore. A universal procedure is described how swelling pressure equilibria can be identified, although the crosslinking density is changed. It can be shown that the gradient of the soluble parts, respectively the additional structural changes of the gel by the soluble parts, is reproducible and with it the swelling pressure equilibria. Furthermore the radial distribution of dyed soluble parts inside the gel phase is presented as a function of time during a sedimentation equilibrium run with a gelatin/water gel. It can be shown that the back diffusion process of the soluble parts in the gel is very slow and does not lead to the sedimentation-diffusion equlibrium of free soluble parts in the considered time interval of 4 months. In combination with the detected fast sedimentation of the soluble component in the gel phase this gives further evidence for the association of the soluble parts to the gel network forming a gradient gel. The predicted formation of the anisotropic gel could be proved by a swelling experiment.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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