Purification and characterization of ATP:AMP phosphotransferase from Mycobacterium marinum

ATP:AMP phosphotransferase (EC 2.7.4.3) (adenylate kinase) has been purified 1746-fold from Mycobacterium marinum (ATCC 927) by successive column chromatography on DEAE-cellulose (DE-53), Reactive Blue agarose, Sephadex G-75, hydroxyapatite and, finally, DEAE-Sephadex A-50. The final enzyme preparation had a specific activity of 576 mumol/min per mg protein with an overall yield of 51%. The preparation was homogeneous by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme was estimated to have an Mr of 29500 and an isoelectric point of 6.7, properties which generally resemble those of the mitochondrial enzyme. Indeed, the two enzymes failed to separate when subjected to polyacrylamide gel electrophoresis under denaturing conditions. The extinction coefficient (at 276 nm) was calculated to be 3.114 X 10(4) M-1 X cm-1 and E1%1cm = 10.556. Adenylate kinase was present at a concentration of 0.06 mg/g (wet weight) bacteria. Enzyme was stable for months in 60% glycerol in the freezer; at 4 degrees C, less than 5% of the activity was lost over a 7 day period.     

Kinetic aspects of the interaction of horse heart cytochrome c with sodium dodecyl sulfate

The sodium dodecyl sulfate (SDS) concentration dependence of spectral changes in circular dichroism (CD) and in absorbance of cytochrome c were examined in the far-ultraviolet region, aromatic region, and the Soret band. The Soret peak obtained in 0.60 mM SDS was nine times greater than that of the native state. (The critical micelle concentration, CMC, of SDS was 2.2 mM in the phosphate buffer used.) The results indicated that the drastic change at the Soret band did not accompany the corresponding large-scale change in secondary structure of the protein. In the stopped-flow measurements, two and three processes were followed at 406 nm below and above the CMC, respectively. At 289 nm only one process was observed, and this corresponded to the second process at 406 nm. Therefore, the second process at 406 nm was considered to be a change in tertiary structure around the heme group. The first process and the third process seemed to reflect a change in the heme environment; the former appeared to be due to a solvent effect and the latter due to a binding effect of a large number of dodecyl sulfate ions.     

THE SPECIES OF CIBORINIA PATHOGENIC TO HERBACEOUS ANGIOSPERMS

Batra , Lekh R., and Richard P. Korf . (Cornell U., Ithaca, N. Y.) The species of Ciborinia pathogenic to herbaceous angiosperms. Amer. Jour. Bot. 46(6): 441–450. Illus. 1959.—On the basis of anatomical studies, 2 distinct species of Ciborinia, a generic segregate of Sclerotinia, are shown to occur on Erythronium. The life history and phytopathology of one of these, C. erythronii, are reported. The species is shown to be homothallic, to develop its sclerotia in nearly all parts of the plant, to overwinter also in the form of mycelium in bulbs, and to form spermatia in the field. Experiments on factors affecting formation of sclerotia and apothecia in the laboratory are reported. Prolonged washing of sclerotia formed in the laboratory proved to be an important factor for development of apothecia. A new species, C. trillii, pathogenic to Trillium is described, as is a new species, C. violae, pathogenic to Viola. The generic diagnosis of Ciborinia is emended on the basis of these studies. A new combination, C. hirtella, is also proposed.     

Nesting behavior ofHalictus scabiosæ in Switzerland (Hymenoptera,Halictidæ)

Summary The structure and populations of seven nests ofHalictus scabiosæ were studied in Geneva during July and August, 1964. Cells, containing brood of various ages, were radially arranged along the branching burrows. The one to six females in each nest did not belong to distinct worker and queen castes. This is in contrast to reports of distinct castes in what appears to be the same species from France. Most pollen collectors in the Geneva population were inseminated and some probably laid eggs.

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Zusammenfassung Die Struktur und die Bewohner von sieben Nestern vonHalictus scabiosæ, die im Juli und August 1964 in der Gegend um Genf gesammelt wurden, werden beschrieben. Zellen mit Brut verschiedenen Alters sind entlang verzweigter Gänge radiär angeordnet. Bis zu sechs Weibchen befanden sich in einem einzelnen Nest. Sie waren nicht in Königinnen und Arbeiterinnen differenziert, wie dies von Bienen aus Frankreich berichtet ist, die wahrscheinlich zur selben Art gehören. Die meisten Pollen-Sammlerinnen der Genfer Population waren befruchtet und einige von ihnen legten auch wahrscheinlich Eier.

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Résumé La structure et la population de sept nids deHalictus scabiosæ ont été étudiées à Genève durant les mois de juillet et d'août 1964. Des cellules contenant du couvain d'âge différent sont disposées en rayons le long de terriers qui se ramifient. Chaque nid était occupé par une à six femelles qui n'appartenaient pas à une caste distincte d'ouvrières ou de reine. La plupart des abeilles qui collectent le pollen furent inséminées et quelques-unes probablement pondèrent des ufs.

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Contribution No. 1303 from the Department of Entomology, The University of Kansas, Lawrence.