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The gene encoding the human erythrocyte form of cytochrome b5 (97 residues in length) has been prepared by mutagenesis of an expression vector encoding lipase-solubilized bovine liver microsomal cytochrome b5 (93 residues in length) (Funk et al., 1990). Efficient expression of this gene in Escherichia coli has provided the first opportunity to obtain this protein in quantities sufficient for physical and functional characterization. Comparison of the erythrocytic cytochrome with the trypsin-solubilized bovine liver cytochrome b5 by potentiometric titration indicates that the principal electrostatic difference between the two proteins results from two additional His residues present in the human erythrocytic protein. The midpoint reduction potential of this protein determined by direct electrochemistry is -9 +/- 2 mV vs SHE at pH 7.0 (mu = 0.10 M, 25.0 degrees C), and this value varies with pH in a fashion that is consistent with the presence of a single ionizable group that changes pKa from 6.0 +/- 0.1 in the ferricytochrome to 6.3 +/- 0.1 in the ferrocytochrome with delta H degrees = -3.2 +/- 0.1 kcal/mol and delta S degrees = -11.5 +/- 0.3 eu (pH 7.0, mu = 0.10). The 1D 1H NMR spectrum of the erythrocytic ferricytochrome indicates that 90% of the protein binds heme in the "major" orientation and 10% of the protein binds heme in the "minor" orientation (pH 7.0, 25 degrees C) with delta H degrees = -2.9 +/- 0.3 kcal/mol and delta S degrees = -5.4 +/- 0.9 eu for this equilibrium. 相似文献
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DV Beard PL Molina KE Muller KM Denelsbeck BM Hemminger JR Perry MP Braeuning DH Glueck WD Bidgood M Mauro 《Canadian Metallurgical Quarterly》1995,197(3):753-758
PURPOSE: Interpretation time of serial staging chest CT cases, which each contained current and previous examinations, with a simple prototype workstation called filmstack was experimentally compared with interpretation time with a film alternator. MATERIALS AND METHODS: The filmstack displayed a "stack" of sections for each examination; user controls allowed rapid selection of preset attenuation windows and both synchronized and unsynchronized scrolling. Eight radiologists were timed as they used the filmstack and the film alternator to interpret four ergonomically complex serial CT cases. RESULTS: All reports dictated on the basis of findings with filmstack and film were of acceptable clinical accuracy. The time to examine a case with filmstack was significantly faster than the time with film, including the time to load and unload the alternator (99% confidence [P = .01]). There was no statistically significant difference in interpretation time between filmstack and prehung film. CONCLUSION: Use of a low-cost stacked CT workstation with a single 1,024 x 1,024 monitor is an effective means of interpreting cases that require comparison of multiple CT examinations. 相似文献
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JM Warnecke JP Fürste WD Hardt VA Erdmann RK Hartmann 《Canadian Metallurgical Quarterly》1996,93(17):8924-8928
To study the cleavage mechanism of bacterial Nase P RNA, we have synthesized precursor tRNA substrates carrying a single Rp- or Sp-phosphorothioate modification at the RNase P cleavage site. Both the Sp- and the Rp-diastereomer reduced the rate of processing by Escherichia coli RNase P RNA at least 1000-fold under conditions where the chemical step is rate-limiting. The Rp-modification had no effect and the Sp-modification had a moderate effect on precursor tRNA ground state binding to RNase P RNA. Processing of the Rp-diastereomeric substrate was largely restored in the presence of the "thiophilic" Cd2+ as the only divalent metal ion, demonstrating direct metal ion coordination to the (pro)-Rp substituent at the cleavage site and arguing against a specific role for Mg(2+)-ions at the pro-Sp oxygen. For the Rp-diastereomeric substrate, Hill plot analysis revealed a cooperative dependence upon [Cd2+] of nH = 1.8, consistent with a two-metal ion mechanism. In the presence of the Sp-modification, neither Mn2+ nor Cd2+ was able to restore detectable cleavage at the canonical site. Instead, the ribozyme promotes cleavage at the neighboring unmodified phosphodiester with low efficiency. Dramatic inhibition of the chemical step by both the Rp- and Sp-phosphorothioate modification is unprecedented among known ribozymes and points to unique features of transition state geometry in the RNase P RNA-catalyzed reaction. 相似文献
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Michael Fellows Jörg Flum Danny Hermelin Moritz Müller Frances Rosamond 《Theory of Computing Systems》2010,46(2):311-339
The classes of the W-hierarchy are the most important classes of intractable problems in parameterized complexity. These classes
were originally defined via the weighted satisfiability problem for Boolean circuits. Here, besides the Boolean connectives
we consider connectives such as majority, not-all-equal, and unique. For example, a gate labelled by the majority connective outputs true if more than half of its inputs are true. For any finite set
C\mathcal{C}
of connectives we construct the corresponding W(
C\mathcal{C}
)-hierarchy. We derive some general conditions which guarantee that the W-hierarchy and the W(
C\mathcal{C}
)-hierarchy coincide levelwise. If
C\mathcal{C}
only contains the majority connective then the first levels of the hierarchies coincide. We use this to show that a variant
of the parameterized vertex cover problem, the majority vertex cover problem, is W[1]-complete. 相似文献
28.
P Mooney M Sarmiento JM Bishop N Biswas NM Cason L Dauwe J Godfrey VP Kenney R Pemper E Rojek RC Ruchti WD Shephard G Ginther RM Edelstein CP Forsyth K Gamarnik AE Kreymer RJ Lipton JM McQuade DM Potter JS Russ L Spiegel DE Johnson D Buchholz LM Cremaldi SW Delchamps HS Mao JL Rosen W Sakumoto RA Schluter SB Sontz C Winter 《Canadian Metallurgical Quarterly》1989,39(9):2494-2498
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