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  • 1
    ISSN: 1573-4978
    Keywords: calf thymus chromatin ; chromatin high order structure ; fiber length ; X-ray small angle scattering
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract This work investigates the structure of native calf thymus chromatin as a function of fiber length and isolation procedures by using X-ray small angle scattering technique. Two methods of chromatin isolation have been compared in order to better understand the differences reported by various authors in terms of chromatin high order structure. In addition to these experimental results the effects of shearing have also been studied. In order to explain the differences among these chromatin preparations we built several models of chromatin fibers (represented as a chain of spherical subunits) assuming increasing level of condensation at increasing salt concentrations. For all these fiber models the corresponding theoretical X-ray scattering curves have been calculated and these results have been used to explain the influence of fiber length on the scattering profiles of chromatin. The comparison between experimental and theoretical curves confirms that the high molecular weight chromatin-DNA prepared by hypotonic swelling of nuclei (without enzymatic digestion) displays a partially folded structure even at low ionic strength, whereas the low molecular weight chromatin-DNA prepared by a brief nuclease digestion appears very weakly folded at the same ionic conditions.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 0006-3525
    Keywords: fourier transform ir spectroscopy ; protein conformations ; cytochrome C ; Langmuir-Blodgett film ; Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: A qualitative and quantitative analysis of the conformation of Langmuir-Blodgett (LB) dried films of cytochrome C on silicon wafers was performed by Fourier transform ir (FTIR) spectroscopy. A deconvolution procedure was applied to the amide I band analysis, in order to determine the percentage of the different secondary structures. Qualitative analysis was performed by examining difference spectra.Films obtained by spreading protein solutions at pH 7.4 and 1, dried at 25 and 100°C, on silicon wafers were also examined in order to detect spectral components associated with denatured protein domains, and to compare them with cytochrome C LB films.FTIR spectroscopy showed that the following important changes characterise LB film spectra: (a) the α-helix component is higher (its percentage is 57 and 54%) than the one estimated in dried film obtained by spreading the solutions at pH 7.4 on a silicon substrate (43%), (b) there is an increase in the intensity of bands attributed to protonated carboxy group bands, involved and not involved in the formation of hydrogen bonds, and a decrease in those attributed to deprotonated carboxy groups, (c) the intensity of several bands attributed to aromatic amino acids and aliphatic chains increases, and (d) bands due to O(SINGLEBOND)H stretching vibrations of crystallization water are present.These conformational changes could be induced by protein-protein interaction caused by the close packing of molecules that occurs during LB film formation; it cannot be excluded that they may be accompanied by partial changes in the tertiary structure of the protein. A preferential orientation of protein molecules in LB films is also a possibility. © 1997 John Wiley & Sons, Inc. Biopoly 42: 227-237, 1997
    Additional Material: 4 Ill.
    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: Theoretical predictions within the Born approximation of the expected differential light scattering of circularly polarized light (CIDS) were made for the 300-Å chromatin fiber, modeled as a helical array of dielectric ellipsoids. Computed CIDS values were strongly dependent on the exact geometry of the solenoid model, depending particularly on parameters relaed to the chiral nature of the fiber and the orientation of the nucleosomes within the helix, in contrast to the values of the total light scattering, which mainly probed size and shape. In particular, both a superbead model and a strict linear 110-Å nucleosome filament would be predicted as giving rise to zero CIDS (in disagreement with the finite values observed). At the same time, helical models in which the normal vectors to the nucleosome faces were exactly parallel to the helical axis also yielded zero CIDS. Confirming earlier expectations, CIDS values were significantly less dependent on helical length than total light scattering. Finally, comparison of these calculated results from those extrapolated from available experimental data indicates that predicted CIDS values, based on currently accepted models of solenoid structure, are within an order of magnitude of those experimentally observed. Together, these results indicate the potential of differential light scattering measurements as a probe of chromatin higher-order structure, complementary to existing scattering measurements.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
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