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51.
The ubiquinone protein, QP-C, in reduced ubiquinone-cytochrome c reductase (the b?c1-III complex) shows a stable ubisemiquinone radical when the enzyme is reduced by succinate in the presence of catalytic amounts of succinate dehydrogenase and QP-S. At room temperature using EPR technique the redox titration of the b?c1-III complex in the presence of redox dyes or succinate/fumarate couple reveals that the ubisemiquinone radical has a midpoint potential of approximately +67 mV at pH 8.0. Further analysis yields E1 of +83 mV and E2 of +51 mV corresponding to () and () or other electronated forms, respectively. The equilibrium radical concentration has been found to be affected both by pH and succinate/fumarate couple. At pH 9.0 the radical shows the maximal amplitude and stability. Below pH 7.0, little radical was detected. The electron spin relaxation behavior of ubisemiquinone radical, as examined by microwave power saturation, indicates that the ubisemiquinone radical of QP-C is somewhat isolated from other paramagnetic centers. The effects of phospholipids, QP-S, and other agents on ubisemiquinone radical formation as well as the enzymatic activity of QP-C have been studied in detail. 相似文献
52.
Controlled alkaline hydrolysis of 16α-bromo-17-keto steroids , and with potassium carbonate and tetra-n-butylammonium hydroxide (n-Bu4NOH) and synthesis of 2α-hydroxy-3-ones , and by the controlled hydrolysis of the corresponding 2α-bromo-3-ones , and are described. Treatm carbonate in aqueous acetone or with n-Bu4NOH in aqueous dimethylformamide (DMF) gave 16α-hydroxy-17-ones , and in 85–90% yield, respectively. 2α-Hydroxy-3-ones , and were obtained by hydrolysis of the corresponding bromoketones , and in high yields using the above conditions or sodium hydroxide in pyridine or DMF, respectively. Deuterium labeling experiments suggested that equilibration between the 2α-bromoketone and the 2β-bromo isomer precedes the formation of the ketol in which the true intermediate might be the 2β-isomer . However, rearranged androstane derivatives, 3β-hydroxy-2-ones and , were stereoselectively obtained by treatment of the bromoketones and with an excess amount of sodium hydroxide. 相似文献
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C A Yu S Nagaoka L Yu T E King 《Biochemical and biophysical research communications》1978,82(4):1070-1078
A highly purified cytochrome complex which is free of QPs (see BBRC , 939) yields 20–30% of semiubiquinone (based on the total Q content in the system) in the presence of catalytic amounts of QPs and succinate dehydrogenase (at or lower than 2 × 10?9 M. The radical shows typical signals with line widths of 8 and 9 gauss, respectively, at about 22°C and 77°K. The appearance of the radicals approximately parallels that of reduction but not its disappearance. However, addition of theonytrifluoroacetone or antimycin A immediately abolishes both radical formation and reduction. These and other observations indicate that the true carrier property of Q is through its binding with proper proteins but not the protei-free form. 相似文献
55.
Mutant 170 not capable of forming streptidine and streptomycin was obtained using chemical mutagenes. This mutant can produce streptomycin only with suplementation of exogenous streptidine. Experiment with labeled C14-streptidine showed its specific incorporation in streptidine moiety of streptomycin molecule. 相似文献
56.
Polynucleotide sequence relationships among Ent plasmids and the relationship between Ent and other plasmids. 总被引:31,自引:13,他引:18 下载免费PDF全文
Deoxyribonucleic acid-deoxyribonucleic acid hybridization studies reveal that the plasmids coding for the production of heat stable and heat labile enteroxtoxins of Escherichia coli, regardless of their origin, have a majority of their polynucleotide sequences in common, but are not related in any significant way to those plasmids coding for the synthesis of only ST toxin. The heat stable and heat labile plasmids also share a significant degree of their polynucleotide sequences with plasmids of the FI and FII incompatibility groups, but not with R factors belonging to the I, N, W, P, or X incompatibility groups. 相似文献
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Molecular cloning of an Escherichia coli plasmid determinant than encodes for the production of heat-stable enterotoxin. 总被引:14,自引:2,他引:12 下载免费PDF全文
A conjugative plasmid, ESF0041 was isolated from an enterotoxigenic strain of Escherichia coli from calves. ESF0041 was found to be 65 x 10(6) daltons in mass of a member of the F incompatibility complex. Acquisition of ESF0041 by E. coli K-12 was invariably associated with the capacity to produce heat-stable (ST) enterotoxin. ESF0041 and pSC101 deoxyribonucleic acids were cleaved with EcoRI, and the fragments were ligated with polynucleotide ligase. Transformation of E. coli K-12 with the ligation mixture led to the isolation of an ST+ clone. Further analysis of the plasmid deoxyribonucleic acid from this clone showed that a structural gene(s) associated with ST biosynthesis had been isolated as a 5.7 x 10(6)-dalton ESF0041 fragment in pSC101. In turn, 5.7 x 10(6)-dalton fragment was ligated to a multicopy COLE1 derivative, RSF2124, so that toxin synthesis was amplified about threefold. 相似文献
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