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1.
Involvement of a protein with molybdenum cofactor in the in vitro activation of nitrate reductase from a chlA mutant of Escherichia coli K12 总被引:1,自引:0,他引:1
Chlorate-resistant mutants are pleiotropically defective in molybdoenzyme activities. The inactive derivative of the molybdoenzyme, respiratory nitrate reductase (nitrite: (acceptor) oxidoreductase, EC 1.7.99.4), which is present in cell-free extracts of chlA mutants can be activated by addition of purified protein PA, the presumed active product of the chlA+ locus, but the activity of the purified protein PA is low, since comparatively large amounts of protein PA are required for the activation. Addition of 10 mM tungstate to the growth medium of a chlBchlC double mutant leads to inactivation of both the molybdenum cofactor and protein PA. Protein PA prepared from such cells was unable to potentiate the in vitro activation of nitrate reductase present in the soluble fraction of a chlA mutant. Quantitation of inactive protein PA was determined immunologically using protein PA-specific antiserum. When a heat-treated extract of a wild-type strain was added to purified protein PA or to the supernatant fraction of a chlBchlC double mutant grown with tungstate, a large stimulation in the ability of these preparations to activate chlA nitrate reductase was found. We equate the activator of protein PA with molybdenum cofactor because: (1) both are absent from heated extracts of tungstate-grown chlBchlC double mutant and cofactor defective chlA and chlE mutants; (2) both are present in heated extracts of wild-type strain; and (3) they behave identically on molecular-sieve columns. 相似文献
2.
Structure and function of hemoglobin in antarctic fishes and evolutionary implications 总被引:2,自引:0,他引:2
Guido di Prisco Rossana D'Avino Laura Camardella Carla Caruso Mario Romano Bruno Rutigliano 《Polar Biology》1990,10(4):269-274
Summary The hematological features of cold-adapted, red-blooded Antarctic teleosts has prompted this study on the relationship between hemoglobin molecular structure and oxygen-binding properties. The hemolysates from 21 species of 5 families contained one component (Hb 1), often accompanied by an additional, minor one (Hb 2, 5%–10% of total). On the other hand, 3 species of Zoarcidae, a non-endemic family, had 4–5 components. All purified hemoglobins from the former group, but only 1–2 of the 4–5 hemoglobins of Zoarcidae, showed a strong Root effect (pH regulation of oxygen binding). Globins from each hemoglobin have been purified and characterised with respect to molecular structure in several species. The similarity between the complete amino acid sequence of one -chain and those of non-Antarctic -chains is lower than that among the latter sequences, suggesting independent pathways of evolution.Presented at the 5th SCAR Symposium on Antarctic Biology, Hobart, Australia (August 29th-September 3rd, 1988) 相似文献
3.
Summary The complete amino acid sequence of the single haemoglobin of the Antarctic fish Aethotaxis mitopteryx DeWitt has been established by automated repetitive Edman degradation on the intact and cleaved (enzymatically and chemically) and chains. A very high sequence identity with other Antarctic fish haemoglobins has been detected. The haemoglobin has a moderate Bohr effect and no Root effect. Organic phosphates and chloride also regulate oxygen binding only to a moderate extent. The lack of Root effect is consistent with the substitution His — Val at the HC3 C-terminal position of the chain. The low overall heat of oxygenation suggests that in this species oxygen transport is an energy-saving process, presumably related to cold adaptation. The comparative analysis of the haemoglobins of Antarctic fishes emphasises some unique features of the oxygen-transport system of A. mitopteryx, which are likely to be related to its also rather unique mode of life.Data presented here were collected during the European Polarstern Study (EPOS) sponsored by the European Science Foundation 相似文献
4.
Ester Cecere Orestina D. Saracino Margherita Fanelli Antonella Petrocelli 《Journal of applied phycology》1992,4(4):323-327
A survey is reported of the drifting algal community in Mar Piccolo, a polluted basin subject to sewage outlets. The key role was played by a few key species, mainly floridean red algae. 相似文献
5.
6.
The chlorate-resistant (chlR) mutants are pleiotropically defective in molybdoenzyme activity. The inactive derivative of the molybdoenzyme, respiratory nitrate reductase, present in the cell-free extract of a chlB mutant, can be activated by the addition of protein FA, the probable active product of the chlB locus. Protein FA addition, however, cannot bring about the activation if 10 mM sodium tungstate is included in the culture medium for the chlB strain. The inclusion of a heat-treated preparation of a wild-type or chlB strain prepared after growth in the absence of tungstate, restores the protein-FA-dependent activation of nitrate reductase. All attempts to activate nitrate reductase in extracts prepared from tungstate-grown wild-type Escherichia coli strains failed. It appears that during growth with tungstate, the possession of the active chlB gene product leads to the synthesis of a nitrate reductase derivative which is distinct from that present in the tungstate-grown chlB mutant. Heat-treated preparations from chlA and chlE mutants which do not possess molybdenum cofactor activity fail to restore the activation. Fractionation by gel filtration of the heat-treated preparation from a wild-type strain produced two active peaks in the eluate of approximate Mr 12000 and less than or equal to 1500. The active material in the heat-treated extract was resistant to exposure to proteinases, but after such treatment the active component, previously of approximate Mr 12000, eluted from the gel filtration column with the material of Mr less than or equal to 1500. The active material is therefore of low molecular mass and can exist either in a protein-bound form or in an apparently free state. Molybdenum cofactor activity, assayed by the complementation of the apoprotein of NADPH:nitrate oxidoreductase in an extract of the nit-1 mutant of Neurospora crassa, gave a profile following gel filtration similar to that of the ability to restore respiratory nitrate reductase activity to the tungstate-grown chlB mutant soluble fraction. This was the case even after proteinase treatment of the heat-stable fraction. Analysis of the chlC (narC) mutant, defective in the structural gene for nitrate reductase, revealed that heat treatment is not necessary for the expression of the active component. Furthermore both the active component and molybdenum cofactor activity are present in corresponding bound and free fractions in the non-heat-treated soluble subcellular fraction.(ABSTRACT TRUNCATED AT 400 WORDS) 相似文献
7.
Maura Turriani Rossana Pesi Antonella Nardone Gino Turchi Francesco Sgarrella Pier Luigi Ipata Maria Grazia Tozzi 《Journal of biochemical and molecular toxicology》1994,9(1):51-57
Nucleoside phosphotransferase acting on inosine and deoxyinosine has been partially purified from cultured Chinese hamster lung fibroblasts (V79). The activity is associated with a cytosolic 5′-nucleotidase acting on IMP and deoxyIMP. The transfer of the phosphate group from IMP to inosine catalyzed by this enzyme was activated by ATP and 2,3-bisphosphoglycerate. Inosine, deoxyinosine, guanosine, deoxyguanosine, and the nucleoside analogs 2′,3′-dideoxyinosine and 8-azaguanosine are substrates, while adenosine and deoxyadenosine are not. IMP, deoxyIMP, GMP, and deoxyGMP are the best phosphate donors. The cytosolic 5′-nucleotidase/phosphotransferase substrate, 8-azaguanosine, was found to be very toxic for cultured fibroblasts (LD50 = 0.32 μM). Mutants resistant to either 8-azaguanosine and the correspondent base 8-azaguanine were isolated and characterized. Our results indicated that the 8-azaguanosine-resistant cells were lacking both cytosolic 5′-nucleotidase and hypoxanthine-guanine phosphoribosyltransferase, while 8-azaguanine resistant cells were lacking only the latter enzyme. Despite this observation, both mutants displayed 8-azaguanosine resistance, thus indicating that cytosolic 5′-nucleotidase is not essential for the activation of this nucleoside analog. 相似文献
8.
Stefano Corsano Giovannella Strappaghetti Rossana Scapicchi Olimpia Scalise 《Bioorganic & medicinal chemistry letters》1993,3(12):2713-2716
The synthesis and evaluaton of the biological activity of a series of 3(2H)-pyridazinones is reported. The bronchodilatating activity of these compounds was determined on the guinea pig trachea. Compounds 6 and 17 with 1-(4-amino-3,5-dichlorophenyl)-2-hydroxyethyl-and 1-(3,4-dichlorophenyl)-2-hydroxyethyl groups linked through a piperazine ring to the 6-position of 3(2H)-pyridazinone show a good bronchospasmolytic activity. 相似文献
9.
Cloning and molecular characterization of two genes encoding adhesion proteins involved in Trichomonas vaginalis cytoadherence 总被引:11,自引:2,他引:9
10.
Signalling of Trichomonas vaginalis for amoeboid transformation and adhesin synthesis follows cytoadherence 总被引:2,自引:0,他引:2
Rossana Arroyo Arturo González-Robles Adolfo Martínez-Palomo J. F. Alderete 《Molecular microbiology》1993,7(2):299-309
The cytoadherence of Trichomonas vaginalis, the sexually transmitted flagellated protozoan, to vaginal epithelial cells (VECs) is the key to infection. Electron microscopy revealed that in vitro-grown parasites having typical globular shape transformed rapidly after contact with VECs into thin, flat, amoeboid cells, maximizing the area of adhesion to the surface of VECs. Amoebic trichomonads formed filopodia and pseudopodia, which interdigitated at distinct sites on the plasma membrane of target cells. In contrast, the amoeboid transformation did not occur for T. vaginalis interacting with He La cells, the previously used in vitro host model cell. Initial parasitism of VECs by a single organism was followed by establishment of a monolayer of trichomonads on the host cell. Finally, parasites adhering to either VECs or HeLa cells were induced to synthesize greater amounts of the four previously described adhesins. Therefore, distinct signals after contact with either epithelial cell type leads to the morphological transformation and/or induction of adhesin synthesis by T. vaginalis. 相似文献