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51.
A ouabain p-aminobenzenediazonium derivative with a high specific radioactivity has been synthesized from ouabain and used as a photolabel for the (sodium plus potassium)-activated adenosinetriphosphatase from Electrophorus electricus electric organ and from dog kidney. In the dark it binds reversibly to the digitalis receptor site, with binding characteristics comparable to those of ouabain. The photoactivation of the ouabain derivative to produced covalent labeling of the receptor was obtained by energy transfer from a tryptophan residue in the (Na+,K+)ATPase to the ouabain p-aminobenzenediazonium molecule bound at the active site. The great advantage of this procedure compared to previous methods is that free molecules of the photoactivatable derivative are not photodecomposed. Analysis of the photolabeled polypeptides on sodium dodecyl sulfate gel electrophoresis showed that over 90% of the total radioactivity incorporated was found in the large molecular weight alpha-chain of the kidney enzyme (Mr 93 000). The same specific labeling of the alpha-subunit was obtained with a crude microsomal fraction from Electrophorus electricus. A mild tryptic fragmentation of the subunit into two peptide fragments of Mr 58 000 and 41 000, respectively, shows that the digitalis receptor is located in the N-terminal 41 000 fragment. 相似文献
52.
Purification and properties of two lectins from the latex of the euphorbiaceous plants Hura crepitans L. (sand-box tree) and Euphorbia characias L. (Mediterranean spurge).
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L Barbieri A Falasca C Franceschi F Licastro C A Rossi F Stirpe 《The Biochemical journal》1983,215(3):433-439
1. From the latex of two members of the plant family Euphorbiaceae, Hura crepitans L. (sand-box tree) and Euphorbia characias L. (Mediterranean spurge), two lectins were purified by affinity chromatography on acid-treated Sepharose 6B followed by elution with D-galactose. 2. The lectin from E. characias is a single molecular species with Mr 80 000, made up of two identical subunits with Mr 40 000, and is a glycoprotein containing 11% carbohydrate. 3. The lectin from H. creptians appears as a mixture of three isolectins with Mr 140 000, consisting of four different subunits with Mr values 37 500, 35 500, 31 000, and 29 000. 4. Both lectins have haemagglutinating activity, with no specificity for human blood groups. The haemagglutinating activity is inhibited by D-galactose and by galactose-containing oligosaccharides. 5. The lectin from H. crepitans is mitogenic to human T-, but not to B-, lymphocytes. The latex of E. characias is mitogenic to T- and, to a lesser extent, to B-, lymphocytes, but the purified E. characias lectin has no mitogenic activity. 6. The lectin from H. crepitans, but not that from E. characias, inhibits protein synthesis by a rabbit reticulocyte lysate. 相似文献
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Differential Recovery of Auxotrophs After Penicillin Enrichment in Escherichia coli 总被引:6,自引:3,他引:3
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Various auxotrophs are recovered from a penicillin enrichment cycle with differing efficiencies. Reconstruction experiments indicate that, under starvation conditions in the presence of penicillin, most auxotrophs undergo some death, whereas prolineless mutants are virtually immune to penicillin-induced killing. 相似文献
56.
Immunoglobulins in human schistosomiasis mansoni 总被引:3,自引:0,他引:3
L J Antunes A P Reis J Pellegrino C A Tavares N Katz 《The Journal of parasitology》1971,57(3):539-542
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Schistosoma mansoni: defined system for stepwise transformation of cercaria to schistosomule in vitro 总被引:38,自引:0,他引:38
F J Ramalho-Pinto G Gazzinelli R E Howells T A Mota-Santos E A Figueiredo J Pellegrino 《Experimental parasitology》1974,36(3):360-372
Defined conditions are described for the in vitro production of large numbers of tail-free viable schistosomules. These consist of (1) the centrifugation of cold cercarial suspension and the incubation of the packed cercariae in a minimal volume of medium at 30 C for 40 min to effect tail loss and glandular secretion; (2) the isolation of the bodies by resuspension and sedimentation and (3) the induction of surface changes by incubating the bodies in inactivated serum or a defined tissue culture medium for a further 40 min interval at 37 C with mild agitation.The resultant schistosomules are characterized by the depletion of their penetration gland contents, loss of tail, fluoride and water sensitivities, complement insensitivity, negative “Cercarien-hüllen Reaktion,” and loss of the surface coat as demonstrated by periodic acid-Schiff (PAS) staining and electron microscope observations. 相似文献
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Summary The osmium tetroxide-zinc iodide fixative of Champy-Maillet has been used to study the rat's retina at the electron microscope level. Electron opaque deposits were observed all along the photoreceptor cells and concentrated in the outer segments of rods and cones and in the nerve endings. In the outer segments that deposits are located in the inter and intra disk spaces as well as between the disk and outer membranes. In the outer plexiform layer reactive sites include synaptic vesicles and mitochondria; other minor reactive sites are described in the inner segment and inner plexiform layer.Electron opaque deposits were not seen if potassium iodide substitutes zinc iodide in the fixative. However, if osmium tetroxide-potassium iodide fixed retinae are immersed in osmium tetroxide-zinc iodide the characteristic electron-dense material is evidenced at those same sites. The effect of other several fixatives were studied with a similar double fixation procedure. Our finding points to the histochemical demonstration of an unidentified component (s) of the retina which shows a striking specificity of localization and which is made evident when zinc iodide is used in the Champy-Maillet mixture.This work has been supported by grants of the Consejo Nacional de Investigaciones Cientificas y Técnicas, Argentina and U.S. Air Force AF-AFOSR 67-0963 A.We are greatly indebted to Miss Haydée Agoff and to Mr. Alberto Saenz for their skillful technical assistance. 相似文献
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