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171.
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NolX of Sinorhizobium fredii USDA257, a Type III-Secreted Protein Involved in Host Range Determination, Is Localized in the Infection Threads of Cowpea (Vigna unguiculata [L.] Walp) and Soybean (Glycine max [L.] Merr.) Nodules
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Hari B. Krishnan 《Journal of bacteriology》2002,184(3):831-839
Sinorhizobium fredii USDA257 forms nitrogen-fixing nodules on soybean (Glycine max [L.] Merr.) in a cultivar-specific manner. This strain forms nodules on primitive soybean cultivars but fails to nodulate agronomically improved North American cultivars. Soybean cultivar specificity is regulated by the nolXWBTUV locus, which encodes part of a type III secretion system (TTSS). NolX, a soybean cultivar specificity protein, is secreted by TTSS and shows homology to HrpF of the plant pathogen Xanthomonas campestris pv. vesicatoria. It is not known whether NolX functions at the bacterium-plant interface or acts inside the host cell. Antibodies raised against S. fredii USDA257 NolX were used in immunocytochemical studies to investigate the subcellular localization of this protein. Immunostaining of paraffin-embedded sections of developing soybean and cowpea (Vigna unguiculata [L.] Walp) nodules revealed localization of NolX in the infection threads. Protein A-gold immunocytochemical localization studies utilizing affinity-purified NolX antibodies revealed specific deposition of gold particles in the fibrillar material inside infection threads. Similar immunogold localization studies failed to detect NolX in thin sections of mature soybean and cowpea nodules. The results from this study indicate that NolX is expressed in planta only during the early stages of nodule development. 相似文献
173.
An Empirical Correlation between Secondary Structure Content and Averaged Chemical Shifts in Proteins 总被引:4,自引:0,他引:4
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It is shown that the averaged chemical shift (ACS) of a particular nucleus in the protein backbone empirically correlates well to its secondary structure content (SSC). Chemical shift values of more than 200 proteins obtained from the Biological Magnetic Resonance Bank are used to calculate ACS values, and the SSC is estimated from the corresponding three-dimensional coordinates obtained from the Protein Data Bank. ACS values of 1Hα show the highest correlation to helical and sheet structure content (correlation coefficient of 0.80 and 0.75, respectively); 1HN exhibits less reliability (0.65 for both sheet and helix), whereas such correlations are poor for the heteronuclei. SSC estimated using this correlation shows a good agreement with the conventional chemical shift index-based approach for a set of proteins that only have chemical shift information but no NMR or x-ray determined three-dimensional structure. These results suggest that even chemical shifts averaged over the entire protein retain significant information about the secondary structure. Thus, the correlation between ACS and SSC can be used to estimate secondary structure content and to monitor large-scale secondary structural changes in protein, as in folding studies. 相似文献
174.
A new binding assay for membrane receptor systems has been developed employing an air-driven ultracentrifuge (Beckman Airfuge). The main advantages of this method for measurement of radioligand binding in aqueous medium are (i) the rapidity (30 s) in separating the bound from the unbound fraction, (ii) the small volume (100 μl) of assay medium which permits a relatively small excess of ligand over receptor to be employed, and (iii) the simplicity of manipulations which allows a high degree of replication. The variation in a triplicate set of assays is usually less than 0.5%. By virtue of maintaining equilibrium throughout the assay the present method is especially useful for ligands exhibiting rapid reversibility in binding. Binding of [3H]ouabain to several membrane (Na+, K+)-ATPases and binding of [3H]etorphine to the oplate receptor from brain membranes are discussed here. Also the inhibition of [3H]ouabain binding by Tris is discussed. 相似文献
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B P Gardner F Theocleous J W Watt K R Krishnan 《BMJ (Clinical research ed.)》1985,291(6509):1620-1622
The case histories of the 37 patients of the Mersey Regional Spinal Injuries Centre who were artificially ventilated between 1968 and 1984 were reviewed. The patients and caring relatives were interviewed separately to determine their views. Of 21 patients who were still alive, 18 said that they would wish to be temporarily ventilated again if the need arose, two were undecided, and one said that she would wish to be allowed to die. Sixteen caring relatives said that they were glad that the decision to ventilate had been taken. Those who were not glad were all young mothers. The outcome from any combination of factors that existed before injury could not be predicted except that most patients over age 59 fared badly. Patients with spinal cord damage should be artificially ventilated if required, provided that this can be carried out well and that total emotional, educational, and physical support can be provided and maintained. Whether or not scarce resources should be deployed in this manner requires discussion. 相似文献