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The isolation and purification of the product of the chl B gene of Escherichia coli K 12 from the chl A mutant have been attempted. The purified protein gives a single band in 10 % sodium dodecylsulfate/polyacrylamide gel electrophoresis. The molecular weight is estimated to be 35 000. This protein, that we have named “FA factor”, does not contain any lipid, has a strong tendency to lose its activity by polymerizing but can be kept in an active state when stored in buffer containing NaCl. The addition of purified FA protein to a soluble extract from the chl B mutant strain grown under anaerobiosis in the presence of nitrate initiates the “complementation reaction”, i.e. the reconstitution of the nitrate reductase activity and the formation of particulate material similar to the native membrane with respect to the structure and enzymatic function. FA protein acts both on the rate of reconstitution and on the total amount of reconstituted enzyme. The complementation leads to the reconstitution of nonsedimentable nitrate reductase and to the formation of three types of particles of different buoyant densities (1.10, 1.18 and 1.23) the two lightest of which contain nitrate reductase. It is shown that FA factor is incorporated only into the particles of intermediate density. In vivo, this factor is located in the native membranes of chl A, chl C, chl D and wild-type strains, whatever the growth conditions, aerobiosis or anaerobiosis, and in the presence or absence of nitrate. Protein FA can be released from either of these membranes (native or reconstituted) by removing Mg2+ or by subjecting Kaback's vesicles to mechanical treatments; in the case of 1.18-reconstituted particles and wild-type membranes, the release of FA protein does not exert any effect on the level of the nitrate reductase activity. 相似文献
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Membrane-bound ATPase (EC 3.6.1.3) of Escherichia coli K 12 is released in a soluble form by the mechanical treatments applied to the cells in order to break them. The purification of the soluble enzyme is described. The purified protein gives a single band in 7.5% polyacrylamide gel electrophoresis. The molecular weight is estimated to be 350 000. The enzyme is cold-labile, Mg-2+ dependent, insensitive to inhibition by N, N'-dicyclohexylcarbodiimide and specific for ATP and ADP. Membranes depleted of their ATPase activity by dilution in a buffer of low ionic strength and without Mg-2+ are able to incorporate the purified ATPase only in the presence of 2-6 mM Mg-2+. ATPase binds to particles formed by complementation between supernatant extracts of chl A and chl B mutants. There are three kinds of particles of different buoyant densities (1.10, 1.18 and 1.23); ATPase binds only to the 1.10 and 1.18 particles. The kinetics of incorporation have been studied. ATPase begins to be incorporated into the 1.10 particles after 10 min of incubation up to a maximum at 20 min: from 30 min, ATPase is incorporated only into 1.18 particles and the amount of incorporated ATPase increased in proportion with the peak of 1.18 particles. These kinetics have a hyperbolic pattern. In order to explain the mechanism of assembly involved in complementation, two hypotheses are proposed. 相似文献
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In the course of a freeze-cleave study on intercellular junctions in the regenerating rat liver, we observed an unusual array of intramembranous particles located in regions of contact between endothelial cells lining the hepatic sinusoids. These arrays were characterized by an accumulation of particles which resembled a zonula occludens in their linear deployment but differed in that the contact regions were composed of individual particles which remained separated from each other by regular particle-free intervals. 相似文献
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Value of cane trash in nitrogen nutrition of sugarcane 总被引:1,自引:0,他引:1
The significance of trash containing 0.3 to 0.5% N in the N nutrition of sugarcane (Saccharum hybrid sp.) was investigated in pot- and field experiments using15N-labelled trash. The data obtained from the pot study with 2 silty-clay loams (a Humic Nitosol and a Humic Acrisol) showed
that surface-applied trash (10 tonnes/ha), although ground to pass a 1-mm sieve, contributed less than 10% of N removed by
sugarcane. Uptake of trash N was most active during the initial 6 months of the experiment though at the end of the study
period of 18 months less than 15% of trash N was altogether recovered by sugarcane.
In the absence of fertilizer N in a field study on the Humic Acrisol (C/N ratio 22), unground trash (5 tonnes. ha−1) depressed soil N uptake by sugarcane by immobilizing available soil N. The field study moreover confirmed that the contribution
of trash N in the supply of N to sugarcane is negligible. The value of trash would reside in its capacity to increase over
the long term the organic matter level in the soils. 相似文献
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