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81.
Gu W Jacquamet L Patil DS Wang HX Evans DJ Smith MC Millar M Koch S Eichhorn DM Latimer M Cramer SP 《Journal of inorganic biochemistry》2003,93(1-2):41-51
We have reexamined the Ni EXAFS of oxidized, inactive (as-isolated) and H(2) reduced Desulfovibrio gigas hydrogenase. Better spatial resolution was achieved by analyzing the data over a 50% wider k-range than was previously available. A lower k(min) was obtained using the FEFF code for phase shifts and amplitudes. A higher k(max) was obtained by removing an interfering Cu signal from the raw spectra using multiple energy fluorescence detection. The larger k-range allowed us to better resolve the Ni-S bond lengths and to define more accurately the Ni-O and Ni-Fe bond lengths. We find that as-isolated, hydrogenase has two Ni-S bonds at approximately 2.2 A, but also 1-2 Ni-S bonds in the 2.35+/-0.05 A range. A Ni-O interaction is evident at 1.91 A. The as-isolated Ni-Fe distance cannot be unambiguously determined. Upon H(2) reduction, two short Ni-S bonds persist at approximately 2.2 A, but the remaining Ni-S bonds lengthen to 2.47+/-0.05 A. Good simulations are obtained with a Ni-Fe distance at 2.52 A, in agreement with crystal structures of the reduced enzyme. Although not evident in the crystal structures, an improvement in the fit is obtained by inclusion of one Ni-O interaction at 2.03 A. Implications of these distances for the spin-state of H(2) reduced H(2)ase are discussed. 相似文献
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Quantitative biochemical analysis of microtubule content in normal and transformed 3T3 cells 下载免费PDF全文
Microtubules in normal and transformed BALB 3T3 cells were preserved in a stabilizing medium and measured by a [3H]colchicine-binding tubulin assay, and compared to total cellular tubulin measured under nonstabilizing conditions. Essentially no change in tubulin or microtubule content was seen with changes in cell density or with changes in cellular morphology at various stages of growth of normal or transformed cells or induced by dibutyryl cAMP treatment of transformed cells. Of five cell lines transformed by a variety of agents, four had a significantly higher total tubulin content than untransformed 3T3 cells and all of them had an increased microtubule content. None of the transformed lines had a lower fraction of tubulin recoverable as sedimentable microtubules compared to untransformed cells, and in three of them this fraction was significantly higher. These results establish that microtubules are present in transformed cells to at least the extent (if not greater) than in normal cells but that there are variations in the total amount of tubulin and microtubules as well as the fraction of the total tubulin present as microtubules which are not strictly correlated with transformation or cell morphology. 相似文献
84.
Zhong J Lai Z Groutas CS Wong T Gan X Alliston KR Eichhorn D Hoidal JR Groutas WC 《Bioorganic & medicinal chemistry》2004,12(23):6249-6254
The pathogenesis of a range of human diseases arises from the aberrant activity of proteolytic enzymes. Agents capable of selectively modulating the activity of these enzymes are of potential therapeutic value. Thus, there is a continuing need for the design of scaffolds that can be used in the development of new classes of protease inhibitors. We describe herein the serendipitous discovery of an unexpected rearrangement that leads to the formation of two novel templates that can be used in the design of protease inhibitors. 相似文献
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The reaction of metal ions with nucleic acids can lead to a variety of dramatic effects on nucleic acid structure, e.g., crosslinking
of the polymer strands, degradation to oligomers and monomers, stabilization or destabilization, and the mispairing of bases.
These effects have important implications for genetic information transfer. Metal ions are involved in many aspects of this
transfer; we are presently concerned with the effect of metal ions on the orientation of the active site of RNA polymerase.
Many of the effects of metal ions on nucleic acid structure involve changes in the conformation of the macromolecules. We
have found that conditions that have been used to convert B DNA to Z DNA lead to at least two other conformational changes,
and phase diagrams delineate the realms of stability of each of the forms. We have carried out a number of studies that demonstrate
that the conversion of B to Z DNA is very closely correlated with a substantial decrease in the ability of the DNA to act
as a template for RNA synthesis.
A portion of this paper has been taken from another paper on “Changes of Biological Significance Induced by Metal Ions in
the Structure of Nucleic Acids,” published in Annali dell' lstituto Superiore di Sanita. 相似文献