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The inhibition of squalene-hopene cyclase (SHC) (E.C. 5.4.99.-), an enzyme of bacterial membranes catalyzing the formation
of pentacyclic sterol-like triterpenes, was studied by using different classes of compounds originally developed as inhibitors
of oxidosqualene cyclase (OCS) (E.C. 5.4.99.7), the enzyme of eukaryotes responsible for the formation of tetracyclic precursors
of sterols. The mechanism of cyclization of squalene by SHC, beginning with a protonation of the 2,3 double bond by an acidic
residue of the enzyme, followed by a series of electrophilic additions of the carbocationic intermediates to the double bonds,
is similar to the mechanism of cyclization of 2,3-oxidosqualene by OSC. The inhibitors studied included: (i) analogs of the
carbocationic intermediates formed during cyclization, such as aza-analogs of squalene and 2,3-oxidosqualene; (ii) affinity-labeling
inhibitors bearing a methylidene reactive group; and (iii) vinyldioxidosqualenes and vinylsulfide derivatives of the substrates.
Comparison of the results obtained with the two enzymes, SHC and OSC, showed that many of the most effective inhibitors of
OSC were also able to inhibit SHC, while some derivatives acted as specific inhibitors. Differences could be easily explained
on the basis of the different substrate specificity of the two enzymes. 相似文献
3.
Oliaro-Bosso S Schulz-Gasch T Balliano G Viola F 《Chembiochem : a European journal of chemical biology》2005,6(12):2221-2228
A structural model of Saccharomyces cerevisiae oxidosqualene cyclase (SceOSC) suggests that some residues of the conserved sequence Pro-Ala-Glu-Val-Phe-Gly (residues 524-529) belong to a channel constriction that gives access to the active-site cavity. Starting from the SceOSC C457D mutant, which lacks the cysteine residue next to the catalytic Asp456 residue Cys457 has been replaced but Asp456 is still there, we prepared two further mutants where the wild-type residues Ala525 and Glu526 were individually replaced by cysteine. These mutants, especially E526C, were very sensitive to the thiol-reacting agent dodecyl-maleimide. Moreover, both the specific activity and the thermal stability of E526C were severely reduced. A similar decrease of the enzyme functionality was obtained by replacing Glu526 with alanine, while substitution with the conservative residues aspartate or glutamine did not alter catalytic activity. Molecular modeling of the yeast wild-type OSC and mutants on the template structure of human OSC confirms that the channel constriction is an important aspect of the protein structure and suggests a critical structural role for Glu526. 相似文献
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During the last few years, the proliferation of miniaturised devices with networking capabilities has provided the technological grounds for pervasive networking environments. It is not visionary to foresee a world of pervasive devices embedded in the environment interacting between them, and with those carried by users, via wireless communications. In addition, fostered by the diffusion of small-size, computational-rich mobile devices, the way content is generated, and accessed is changing with respect to the legacy-Internet paradigm. An ever-increasing share of the Internet content is generated directly by the users, and shared on the network (following the User-Generated Content model). While today the legacy Internet is still used to share user-generated content, it is reasonable to envision that pervasive networking technologies will represent the natural platform to support this new model. This will result in content being distributed on users’ devices rather than on centralised servers on the Internet, and in users creating ad hoc networks to share content. The p2p paradigm is particularly suitable for this scenario, because communications will occur directly among users, instead of being necessarily mediated by centralised servers. Motivated by these remarks, in this work we focus on p2p multicast services over ad hoc networks aimed at sharing content among groups of users interested in the same topics. Specifically, starting from a reference solution in legacy wired networks (Scribe), we design a cross-layer optimised protocol (XScribe) that addresses most of the Scribe problems on ad hoc networks. XScribe exploits cross-layer interactions with a proactive routing protocol to manage group membership. Furthermore, it uses a lightweight, structureless approach to deliver data to group members. By jointly using experimental results and analytical models, we show that, with respect to Scribe, XScribe significantly reduces the packet loss and the delay experienced by multicast receivers, and increases the maximum throughput that can be delivered to multicast groups. 相似文献
6.
Shah Ghazanfar Ali Polette Arnaud Pernot Jean-Philippe Giannini Franca Monti Marina 《Engineering with Computers》2021,37(4):2891-2909
Engineering with Computers - This paper introduces a new fitting approach to allow an efficient part-by-part reconstruction or update of editable CAD models fitting the point cloud of a digitized... 相似文献
7.
Osmar Franca Siebel Marcio Cherem Schneider Carlos Galup-Montoro 《Microelectronics Journal》2012,43(5):329-336
This paper exploits a universal current-based definition of the threshold voltage (VT) and discusses some direct methods to measure it. The consistency, accuracy, and sensitivity of the extraction procedures to second-order effects are examined through numerical simulations and experimental measurements. In addition to three procedures based on dc current measurements we propose an automatic VT-extractor circuit which allows the direct determination of the threshold voltage with minimum influence of second-order effects. 相似文献
8.
This paper describes an electrically programmable switched-capacitor (SC) biquad using quasi-passive algorithmic digital-to-analog converters (DAC's). Since only two equal-valued capacitors are needed for programming each capacitance value, the proposed technique offers compact, cost-effective programmability when compared to traditional programming techniques employing binary-weighted capacitor-arrays (C-arrays). A demonstration prototype chip realized in a 1.2 μm CMOS double-metal double-poly technology, and which implements an 8 b programmable SC biquad giving a wide range of lowpass, bandpass and highpass filtering functions, occupies an active area of only 0.38 mm2 相似文献
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10.
The effect of nerve growth factor (NGF) on ontogenesis of frog mast cells was investigated in vivo by histochemical, morphometric, and ultrastructural analysis. Three groups of tadpoles at various stages of development were used. In the first group, the larvae received i.p. injections of 1 ng NGF/g; the second group received 10 ng NGF/g, while the control group received only the vehicle. The first recognizable mast cells arose symmetrically in the tongue at stage 26 of Witschi's standard table. At stages 26 and 29, the mast cell number in the NGF-injected tadpoles was significantly higher than the control group. From stage 29 onward, the mast cell number rapidly increased in all groups. No significant differences in mast cell number were observed between the control group and the NGF-injected groups at stages 31 and 33. Electron microscopy revealed that at metamorphic climax (stage 33), the mast cells in the NGF-treated groups were more mature than those in the control group. Therefore, nerve growth factor at early stages of tadpole development is likely to induce differentiation of mast cell precursors, while at later stages it is likely to induce maturation of immature mast cells. The close anatomical association between mast cells and perineurium, observed during nerve development, is intriguing. Already in the early stages of nerve development, the mast cells form a network around Schwann cell-axon complexes, together with the perineurial cells. At climax, the mast cells are located between the perineurial layers, suggesting that they may play a role in the tissue-nerve barrier of the perineurium. Nerve growth factor also seems to induce perineurial cell maturation. 相似文献