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31.
Karnachuk OV Kurochkina SY Nicomrat D Frank YA Ivasenko DA Phyllipenko EA Tuovinen OH 《Antonie van Leeuwenhoek》2003,83(1):99-106
A sulfate-reducing bacterium, designated as strain R2, was isolated from wastewater of a ball-bearing manufacturing facility
in Tomsk, Western Siberia. This isolate was resistant up to 800 mg Cu/l in the growth medium. By comparison, Cu-resistance
of reference cultures of sulfate-reducing bacteria ranged from 50 to 75 mg Cu/l. Growth experiments with strain R2 showed
that Cu was an essential trace element and, on one hand, enhanced growth at concentrations up to 10 mg/l but, on the other
hand, the growth rate decreased and lag-period extended at copper concentrations of >50 mg/l. Phenotypic characteristics and
a 1078 bp nucleotide sequence of the 16S rDNA placed strain R2 within the genus Desulfovibrio. Desulfovibrio R2 carried at least one plasmid of approximately of 23.1 kbp. A 636 bp fragment ot the pcoR gene of the pco operon that encodes Cu resistance was amplified by PCR from plasmid DNA of strain R2. The pco genes are involved in Cu-resistance in some enteric and aerobic soil bacteria. Desulfovibrio R2 is a prospective strain for bioremediation purposes and for developing a homologous system for transformation of Cu-resistance
in sulfate-reducing bacteria.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
32.
Dimer-tetramer transition between solution and crystalline states of streptavidin and avidin mutants
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Pazy Y Eisenberg-Domovich Y Laitinen OH Kulomaa MS Bayer EA Wilchek M Livnah O 《Journal of bacteriology》2003,185(14):4050-4056
The biotin-binding tetrameric proteins, streptavidin from Streptomyces avidinii and chicken egg white avidin, are excellent models for the study of subunit-subunit interactions of a multimeric protein. Efforts are thus being made to prepare mutated forms of streptavidin and avidin, which would form monomers or dimers, in order to examine their effect on quaternary structure and assembly. In the present communication, we compared the crystal structures of binding site W-->K mutations in streptavidin and avidin. In solution, both mutant proteins are known to form dimers, but upon crystallization, both formed tetramers with the same parameters as the native proteins. All of the intersubunit bonds were conserved, except for the hydrophobic interaction between biotin and the tryptophan that was replaced by lysine. In the crystal structure, the binding site of the mutated apo-avidin contains 3 molecules of structured water instead of the 5 contained in the native protein. The lysine side chain extends in a direction opposite that of the native tryptophan, the void being partially filled by an adjacent lysine residue. Nevertheless, the binding-site conformation observed for the mutant tetramer is an artificial consequence of crystal packing that would not be maintained in the solution-phase dimer. It appears that the dimer-tetramer transition may be concentration dependent, and the interaction among subunits obeys the law of mass action. 相似文献
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Janus (Jak) tyrosine kinases contain a tyrosine kinase (JH1) domain adjacent to a catalytically inactive pseudokinase domain (JH2). The JH2 domain has been implicated in regulation of Jak activity, but its function remains poorly understood. Here, we found that the JH2 domain negatively regulates the activity of Jak2 and Jak3. Deletion of JH2 resulted in increased tyrosine phosphorylation of the Jak2- and Jak3-JH2 deletion mutants as well as of coexpressed STAT5. In cytokine receptor signaling, the deletion of the Jak2- and Jak3-JH2 domains resulted in interferon-gamma and interleukin-2-independent STAT activation, respectively. However, cytokine stimulations did not further induce the JH2 deletion mutant-mediated STAT activation. The deletion of the Jak2 JH2 domain also abolished interferon-gamma-inducible kinase activation, although it did not affect the reciprocal Jak1-Jak2 interaction in 293T cells. Chimeric constructs, where the JH2 domains were swapped between Jak2 and Jak3, retained low basal activity and cytokine inducible signaling, indicating functional conservation between the two JH2 domains. However, the basal activity of Jak2 was significantly lower than that of Jak3, suggesting differences in the regulation of Jak2 and Jak3 activity. In conclusion, we found that the JH2 domain has a conserved function in Jak2 and Jak3. The JH2 domain is required for two distinct functions in cytokine signaling: (i) inhibition of the basal activity of Jak2 and Jak3, and (ii) cytokine-inducible activation of signaling. The Jak-JH2 deletion mutants are catalytically active, activate STAT5, and interact with another Jak kinase, but the JH2 domain is required to connect these signaling events to receptor activation. Thus, we propose that the JH2 domain contributes to both the uninduced and ligand-induced Jak-receptor complex, where it acts as a cytokine-inducible switch to regulate signal transduction. 相似文献
37.
Olsen H Hedengran Faulds MA Saharinen P Silvennoinen O Haldosén LA 《Biochemical and biophysical research communications》2002,293(1):139-144
Ornithine decarboxylase (ODC), the first rate-limiting enzyme in the polyamine biosynthesis is one of the most rapidly degraded proteins in eukaryotic cells. Mammalian ODC is a notable exception to the widely accepted dogma that ubiquitination is always required for targeting a protein to degradation by the 26S proteasome. However, while it is well established that in mammalian cells degradation of ODC is ubiquitin independent, the requirement of ubiquitination for degradation of ODC in yeast cells remained undetermined. We have investigated ODC degradation in three mutant strains of Saccharomyces cerevisiae in which ubiquitin-dependent protein degradation activity is severely compromised. While yeast ODC was rapidly degraded in all these mutant strains the degradation of N-end rule substrates was inhibited. A mutant mouse ODC that fails to interact with Az was rapidly degraded in yeast cells but was stable in mammalian cells suggesting that interaction with a mammalian Az like yeast protein is not necessary for the degradation of ODC in yeast cells. Deletion analysis revealed that sequences from its unique N-terminus are involved in targeting yeast ODC to rapid degradation in yeast cells. 相似文献
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Luodonpää M Rysä J Pikkarainen S Tenhunen O Tokola H Puhakka J Marttila M Vuolteenaho O Ruskoaho H 《Regulatory peptides》2003,111(1-3):153-159
Xenin is a 25 amino acid peptide produced by specific endocrine cells of the duodenal mucosa. Xenin has multiple biological actions in the gastrointestinal tract. It modulates intestinal motility, affects exocrine pancreatic secretion, and gastric secretion of acid. In the present investigation, we studied plasma concentration of xenin in volunteers after modified sham feeding and after meals of different composition. Plasma xenin concentrations were determined by radioimmunoassay in unextracted plasmas and after acidic extraction using C-18 Sep-Pak chromatography and after neutral extraction using affinity filtration. Both extraction methods were followed by C 18 r.p. HPLC chromatography. Xenin plasma concentrations in unextracted and in extracted plasma rose significantly after modified sham feeding when the food was brought to the volunteers from another room immediately before sham feeding started. When the volunteers had the opportunity to observe the preparation of the meal, xenin plasma concentrations during fasting were high and no further rise was observed after sham feeding. Isocaloric feeding resulted in elevated xenin concentrations in unextracted plasma and after high-pressure liquid chromatography. The methods of extraction, acidic or neutral, did not affect the results. CONCLUSION: Cephalic factors, investigated by modified sham feeding, stimulate release of xenin into the circulation. Xenin may participate in the central nervous regulation of gastrointestinal function. 相似文献
40.
The effect of dietary phosphorus deficiency on the immune responses of European whitefish (Coregonus lavaretus L.) 总被引:1,自引:0,他引:1
Low phosphorous (P) feeds in aquaculture are recommended to reduce eutrophication of water systems. However, the feed should be adequate for normal growth and intact immune defence. Influence of low dietary P supply on non-specific and specific immune defence of European whitefish (Coregonus lavaretus) was studied in two trials. For Trial 1, a semi-purified, low-P diet was formulated and supplied with 0, 2.5, 5.0, 7.5, or 10.0 g P kg(-1)to obtain a P gradient of 4.4-14.9 g kg(-1)diet. Diets were fed to four replicate groups of fingerling whitefish for 42 days in a flow-through, freshwater system maintained at 15 degrees C. Fish fed with the P-unsupplemented diet had significantly lower plasma immunoglobulin M (IgM) levels than fish fed with the P-fortified diets (means 2.53 vs. 3.19 mg ml(-1); P=0.047). Plasma lysozyme activity did not differ between fish fed with the P-unsupplemented diet and fish fed with the P-fortified diets (means 21.8 vs. 25.5 U ml(-1); P=0.107). For Trial 2, fish were acclimated for 49 days by feeding the lowest or highest dietary P contents, and thereafter immunised with a single intraperitoneal injection of a novel antigen, bovine gamma globulin (BGG). A clear antibody response against BGG was elicited but the antibody levels did not differ between fish fed low-P or high-P contents. Similar to Trial 1, plasma IgM was lower in fish fed low-P diets, and low-P diet did not influence plasma lysozyme activity. In both trials the growth of fish, having received low-P diet, was significantly lowered. It is concluded that P deficiency has only minor effects on the immune parameters in whitefish, and the practical aquafeed with P contents sufficient for normal growth does not compromise immune functions of this species. 相似文献