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271.
The effect of protamine on growing cells of Escherichia coli K-12 strains containing different K+-uptake systems was investigated. Immediately after the addition of the toxic peptide, growth ceased and all strains lost
most of their K+. In addition, these cells released a significant amount of their ATP into the medium, and the cytoplasmic volume of these
cells decreased by 70%. Whereas cells without rapid K+-uptake systems did not recover, cells containing either the Trk systems or the overproduced Kup system slowly reversed the
effects of protamine, and growth resumed after the cells had reached their original volume. Experiments with a set of strains
carrying mutations in the K+-uptake gene trkA showed a reasonably satisfactory correlation between inhibition of net K+ uptake and the lag time for resumption of growth after addition of protamine. Cells carrying mutations in three extracytoplasmic
proteases were hypersusceptible to protamine, suggesting that the toxic peptide is degraded by these proteases. Data on the
effect of a second addition of protamine suggest that protamine degradation activity is inducible. These data are interpreted
to mean that reaccumulation of K+ by protamine-treated cells triggers recovery of the cells, thereby allowing induction of extracytoplasmic proteases. These,
in turn, degrade protamine, leading to complete recovery of the cells and resumption of growth. Cells that cannot take up
K+ rapidly remain metabolically compromised to such an extent that extracytoplasmic protease activity is not induced, leading
to a prolonged susceptibility of the cells to the toxic peptide.
Received: 27 August 1996 / Accepted: 25 November 1996 相似文献
272.
Difficulties of molecular dissociation of Clostridium botulinum type G progenitor toxin 总被引:1,自引:0,他引:1
Masafumi Nukina Yumi Mochida Sumiko Sakaguchi Genji Sakaguchi 《FEMS microbiology letters》1991,79(2-3):165-170
Clostridium botulinum type G progenitor toxin was chromatographed on DEAE-Sephadex and Q-Sepharose equilibrated with 0.05 M Tris-HCl buffer, pH 8.0, containing 0.2 M urea. The toxin was eluted in a single protein peak from DEAE-Sephadex, but it was eluted in four protein peaks from Q-Sepharose; the third peak was toxic and the others were nontoxic. The third peak, appearing to be the toxic component, had a molecular mass of 150,000. In SDS-polyacrylamide gel electrophoresis, purified type G progenitor toxin migrated in six bands, with molecular masses of 150,000, 140,000, 58,000, 10,800, 10,600, and 10,400. Type G progenitor toxin may be composed of a toxin component with a molecular mass of 150,000 and a nontoxic component in a manner similar to progenitor toxins of other types. Type G toxic component, whether it was reduced or not, migrated in a single band to the same relative positions in SDS-PAGE; type A toxic component reduced with 2-mercaptoethanol migrated in two bands. 相似文献