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Cobalamin-dependent methionine synthase (5-methyltetrahydrofolate-homocysteine methyltransferase, EC 2.1.1.13) has been isolated from Escherichia coli B in homogeneous form. The enzyme is isolated in an inactive form with the visible absorbance properties of cob(II)alamin. The inactive enzyme exhibits an electron paramagnetic resonance (EPR) spectrum at 38 K that is characteristic of cob(II)alamin at acid pH, where the protonated dimethylbenzimidazole substituent is not coordinated with the cobalt nucleus (base-off cobalamin). An additional, variable component of the EPR spectrum of the inactive enzyme has the characteristics of a cob(III)alamin-superoxide complex. Previous work by others [Taylor, R.T., & Weissbach, H. (1969) Arch. Biochem. Biophys. 129, 745-766. Fujii, K., & Huennekens, F.M. (1979) in Biochemical Aspects of Nutrition (Yagi, K., Ed.) pp 173-183, Japan Scientific Societies, Tokyo] has demonstrated that the enzyme can be activated by reductive methylation using adenosylmethionine as the methyl donor. We present data indicating that the conversion of inactive to methylated enzyme is correlated with the disappearance of the EPR spectrum as expected for the conversion of paramagnetic cob(II)alamin to diamagnetic methylcobalamin. When the methyl group is transferred from the methylated enzyme to homocysteine under aerobic conditions, cob(II)alamin/cob(III)alamin-superoxide enzyme is regenerated as indicated by the return of the visible absorbance properties of the initially isolated enzyme and partial return of the EPR spectrum. Our enzyme preparations contain copper in approximately 1:1 stoichiometry with cobalt as determined by atomic absorption spectroscopy.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
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Construction and expression of synthetic DNA fragments coding for polypeptides with elevated levels of essential amino acids 总被引:1,自引:0,他引:1
J. M. Jaynes P. Langridge K. Anderson C. Bond D. Sands C. W. Newman R. Newman 《Applied microbiology and biotechnology》1985,21(3-4):200-205
Summary Polypeptides, with elevated levels of essential amino acids, could be useful as partial protein supplements to food and feeds. To obtain DNA fragments coding for these polymers, oligonucleotides were constructed by random synthesis of a mixture of appropriate codon pairs and inserted into a bacterial plasmid in E. coli. Two of the isolated fragments were subjected to DNA sequence analysis and theoretically code for polypeptides containing up to 23% lysine, 12% tryptophan, 12% methionine, 6% isoleucine, and 6% threonine. These five amino acids make up 60% of the total amino acid content of the peptide, compared with 25% for the same amino acids in lactalbumin, a milk protein considered to be high in essential amino acids. These fragments, when fused to an active bacterial promoter, which directs the synthesis of chloramphenicol acetyl transferase (CAT), cause bacteria, harboring these modified genes, to take up more lysine as compared to control cells and produce commensurately larger CAT polypeptides. This method of gene synthesis may permit production of polypeptides with a specified amino acid composition to supplement specific diets low in the essential amino acids. 相似文献
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Site-related differences in the localization of the monoclonal antibody OX7 in SL2 and SL1 lymphomas
Howard Sands Peter L. Jones William P. Neacy Sudhir A. Shah Brian M. Gallagher 《Cancer immunology, immunotherapy : CII》1986,22(3):169-175
Summary The uptake of a monoclonal antibody (OX7) by murine lymphomas (SL1, SL2) growing in two sites in the mouse were compared. SL2 tumors grown in the subrenal site showed greater specific antibody uptake than did the same tumor grown in the subcutaneous site. Major differences in membrane bound antibody, in vitro antibody binding patterns, and gamma scintillation camera imaging were also observed between the two sites. These differences may be due to the greater blood flow measured in tumors growing in the subrenal capsule than those growing at the subcutaneous site. The differences observed in antibody uptake of the same tumor growing in two different sites raises questions concerning the choice of animal model systems that can be used to predict clinical utility. 相似文献
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Recent spectroscopic and magnetic susceptibility studies of the iron center in the two-iron ferredoxins provide criteria which any model for the iron-sulfur complex in these proteins must satisfy. These criteria are most stringent for parsley and spinach ferredoxin: the reduced proteins contain a high-spin ferric atom antiferromagnetically exchange-coupled (presumably via sulfide bridging ligands) to a high-spin ferrous atom. In the oxidized proteins the iron atoms are antiferromagnetically spin-coupled, high-spin ferric atoms. Arguments are given to substantiate the claim that the ferrous atom in the reduced protein is ligated by four sulfur atoms in a distorted tetrahedral configuration: two are the bridging sulfides, two are cysteinyl sulfurs. A treatment of proton contact shifts based upon the above model is pertinent to proton magnetic resonance data already available and provides a means to identify directly the ligands at both iron atoms via further PMR experiments. 相似文献
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Improved Solid Medium for the Detection and Enumeration of Pectolytic Bacteria 总被引:7,自引:1,他引:6
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An improved solid agar medium (MP medium) has been developed which allows detection of pectolytic activity in bacteria. Organisms tested exhibited a variety of regulatory controls governing pectate lyase synthesis. The medium contains mineral salts, pectin, and yeast extract. After growth of the organisms, the agar plate is flooded with a polysaccharide precipitant, and pectolytic activity is shown by clear zones around active colonies. High concentrations of phosphate are shown to be necessary for pectic enzyme formation on solid media. The medium has successfully been used to detect pectolytic organisms in soil, forest litter, and rotting vegetable samples. 相似文献
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W. A. Sands 《Insectes Sociaux》1965,12(2):117-130
Summary Alates ofTrinervitermes appear in the mounds two or three weeks before their first flights. Those ofT. carbonarius fly first towards the end of May, the other four species in mid-June. Development of alates inT. ebenerianus was accelerated by moisture. The mean sex ratio of all five species in 3664 per cent, males to females, comparable to the percentages of soldiers and workers in young colonies. Flights of alates are late nocturnal, most often at low air temperatures with high relative humidities. «Tandem behaviour» follows wing shedding and precedes «digging-in». A relatively strong stimulus is needed to initiate flight. Thimogtaxis is important in nest site selection. Development of young colonies was followed in glass plate observation nests. The timetable of main events is given for all five species; oviposition and hatching rates and lengthsof instars are tabulated comparatively. The rates of development of young colonies are discussed in relation to their possible adaptive significance.
Résumé On trouve les ailés deTrinervitermes dans leurs termitières de deux à trois semaines avant les premiers vols. ChezT. carbonarius, les vols commencent vers la fin de mai, et chez les autres espèces à la mi-juin. On accélérait la croissance des nymphes deT. ebenerianus en humidifiant les termitières. Le rapport moyen des sexes est 36,64 pour cent, mâles à femelles, et c'est en proportion comparable au rapport entre les soldats et les ouvriers dans les colonies jeunes. Les vols des ailés ont lieu à une heure avancée de la nuit, et ils se présentent le plus souvent en conditions de températures peu élevées, et d'humidités relatives élevées. Le «comportement en tandem» suit l'amputation des ailes et précède le creusement d'une chambre nuptiale. La thygmotaxie est importante en ce qui concerne le choix d'un emplacement pour cette chambre. On a observé le développment des colonies nouvelles dans des nids sous verre. La table chronologique des événements d'importance est donnée pour les cinq espees. On a disposé en tables les données relatives à l'oviposition et à l'éclosion pour effectuer des comparaisons entre les cinq espèces. On examine les vitesses de croissance des jeunes colonies en fonction de leurs significations adaptives.相似文献