The DNA molecule is extremely compacted in bacteria, in cell nuclei, sperm heads and virus capsids. These interactions between DNA molecules are important to our understanding of chromatin condensation. DNA forms multiple liquid-crystalline phases whose nature depends on the polymer concentration, and it has been suggested that the highly concentrated phase of 50-nm DNA molecules is two-dimensionally ordered and smectic-like. We rule out this smectic hypothesis and demonstrate by polarizing microscopy, electron microscopy and X-ray diffraction that this phase is characterized by a columnar longitudinal order and a hexagonal lateral order, with intermolecular distances ranging from 2.8 to 4.0 nm depending on the DNA concentration.
The putative transformation of alpha-helices into beta-sheets has been studied for more than 50 years in the case of hard alpha-keratin. In a previous study of stretched keratin fibers, we specified the conditions for beta-sheet appearance within horsehair: the formation of beta-sheets requires at least 30% relative humidity. However, this phenomenon was observed in the whole tissue. Then there was no clear chemical identification of the beta-sheets (keratin or matrix proteins) and the exact location of the beta-sheets across the fiber could not be specified. In this study, using wide-angle x-ray scattering and high spatial resolution infrared microspectroscopy, we could determine and characterize the structural elements across hair sections stretched in water, which provides new information about the aforementioned transition. Our results show that the process can be split into three steps: 1), unraveling of the alpha-helical coiled-coil domains, which starts at roughly 5% macroscopic strain; 2), further transformation of the unraveled coiled-coils into beta-sheet structures, which occurs above roughly 20% macroscopic strain; and 3), spatial expanding of the beta-structured zones from the sample center to its periphery.
Conditions of formation of DNA aggregates by the addition of spermidine were determined with 146 base pair DNA fragments as a function of spermidine and NaCl concentration. Two different phases of spermidine-DNA complexes are obtained: a cholesteric liquid crystalline phase with a large helical pitch, with interhelix distances ranging from 31.6 to 32.6 A, and a columnar hexagonal phase with a restricted fluidity in which DNA molecules are more closely packed (29.85 +/- 0.05 A). In both phases, the DNA molecule retains its B form. These phases are always observed in equilibrium with the dilute isotropic solution, and their phase diagram is defined for a DNA concentration of 1 mg/ml. DNA liquid crystalline phases induced by spermidine are compared with the DNA mesophases already described in concentrated solutions in the absence of spermidine. We propose that the liquid crystalline character of the spermidine DNA complexes is involved in the stimulation of the functional properties of the DNA reported in numerous experimental articles, and we discuss how the nature of the phase could regulate the degree of activity of the molecule.
In aqueous solution, pure DNA forms multiple liquid crystalline and crystalline phases whose nature depends on the polymer concentration. The following phase sequence is observed when the DNA concentration increases : isotropic ↦ cholesteric ↦ columnar hexagonal ↦ crystalline phases. The aim of this work is to obtain structural information about the highly concentrated phases formed by 500 Å long DNA molecules — in particular about the crystalline phases — by means of X-ray diffraction. We show that in the two-dimensional (2D) ordered hexagonal phase a longitudinal order progressively appears between neighbouring DNA helices leading continuously to a three-dimensional (3D) ordered hexagonal phase. For higher concentrations the specimens undergo a discontinuous transition towards an orthorhombic phase. The characteristic structural parameters of these different phases have been determined. An important result is that the number of nucleotides per helix turn decreases continuously, when the DNA concentration increases, from 10.3±0.1 at the cholesteric ↦ hexagonal transition down to 9±0.1 without any apparent change of the B conformation of the molecules.
Various types of human stratum corneum (sheets or callus) were exposed, in parallel and perpendicular geometry, to the high flux of X rays produced by a synchrotron radiation source. Under these conditions, very clear and rich diffraction patterns, corresponding to the supramolecular organization of stratum corneum proteins and lipids, were obtained. The comparative study of normal or delipidized stratum corneum sheets and membrane couplets allows one to attribute certain diffraction features to lipids. Our results in the 3-7-nm range show two different distances for lipid bilayers. Concerning the protein nature of normal stratum corneum, the results show that keratin would occur in the beta form, whereas for callus it is in the alpha form. Indeed, normal stratum corneum sheets never display the 0.514-nm characteristic of alpha keratin. This result means that the supramolecular organization of keratin could depend on the keratinization process. Finally, our studies also confirm the presence of a still-unknown protein component existing in the beta form that would be located either inside the corneocytes or in some dilatated zones of the intercellular spaces.
We present here a study of the ’’rotator’’ phase displayed by the odd-numbered normal paraffins CnH2n+2 with n ranging from 17 to 21. A structural model of the rotator phase can be deduced from x-ray experiments performed both on single domains and on powder samples. Its structure very much looks like the crystalline phase structure, i.e., the molecules are packed within layers forming a bilayer structure with the molecular axes oriented perpendicularly to the layer planes. The comparison of the space group Ccmm with the steric dimensions of the molecules implies the appearance of an orientational disorder of the molecules around their long axes which is the main characteristic of the crystalline→rotator phase transition. Such a conclusion is in agreement with the dynamical measurements which state a uniaxial rotation of the molecules.
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