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21.
Zusammenfassung Die Abtrennung der OES-Decarboxylase vom Malic-Enzym wird bei Enzymlösung aus Bakterien des Stammes Lactobacillus plantarum L durchgeführt. Die Trennung gelingt wegen der unterschiedlichen Temperatur-und pH-Empfindlichkeit der beiden Enzyme. Das Malic-Enzym ist im Bereich von pH 4,5–5,0 nur dann bei 40° C und darüber vor Denaturierung geschützt, wenn es als Substrat-NAD+-Mn++-Komplex vorliegt. Die OES-Decarboxylase ist entsprechend temperaturstabiler als das Malic-Enzym, auch wenn keine Oxalessigsäure zugesetzt wird.Die Ammoniumsulfat-Fraktionierung bringt keine befriedigende Trennung der Enzyme mit sich. Wird das Malic-Enzym durch Denaturierung nicht entfernt, so reißt es bei 75% iger Sättigung mit Ammoniumsulfat die OES-Decarboxylase mit in den Niederschlag. Bei dieser Salzkonzentration fällt auch die Malat-Dehydrogenase aus.Mikrobiolosisch und papierchromatographisch durchgeführte Biotinnachweise bestätigen, daß die OES-Decarboxylase des Stammes, L ein Biotinproteid ist. 相似文献
22.
Zusammenfassung Die Malat-abbauenden Enzyme von drei L. plantarum-Arten verschiedener Herkunft werden untersucht und miteinander verglichen. Neben Malic-Enzym enthalten alle drei Arten Malat-Dehydrogenase und Oxalessigsäure-Decarboxylase sowie Lactat-Dehydrogenase.Die Oxalessigsäure-Decarboxylase der auf pflanzlichem Material vorkommenden Bakterien (L. plantarum L und L. plantarum ATCC-8014) dürfte nach den Ergebnissen von Hemmversuchen mit Avidin ein Biotin-Proteid sein; beim Malic-Enzym kann dies ausgeschlossen werden.Die Enzymausstattung eines aus der Mundhöhle isolierten Stammes (L. plantarum ATCC-11580) unterscheidet sich insofern von der der übrigen Stämme, als die MDH-Aktivität relativ schwach ausgeprägt ist und das Malic-Enzym ein Biotin-Proteid darstellt. 相似文献
23.
24.
M. Rosa Pinol Urs Kägi Claus W. Heizmann Brigitte Vogel Jean-Marc Séquier Werner Haas Willi Hunziker 《Journal of neurochemistry》1990,54(6):1827-1833
Many processes in the CNS depend on calcium. The calcium signal is transduced into an intracellular response via Ca2(+)-binding proteins, including calbindin D-28K. In many laboratories, polyclonal antibodies against chicken intestinal calbindin D-28K have been used to study its localization in the brain (normal and degenerated) of various species, including humans, but some of these antisera cross-reacted with other proteins, including calretinin. We purified recombinant rat brain calbindin D-28K to raise antisera in rabbits and purified a recombinant rat-chicken calbindin D-28K hybrid protein to immunize mice for the generation of monoclonal antibodies. These antisera were highly specific for calbindin D-28K, as demonstrated by two-dimensional Western blotting analysis. Immunohistochemical analyses combined with in situ hybridization studies demonstrated that calbindin D-28K in the Purkinje cells of the cerebellum is independent of vitamin D. The antibodies described here will be important tools for studying the regulation of expression of calbindin D-28K and its biological function in the brain and in the PNS. 相似文献
25.
Morphological and metabolic characterization of a new species of strictly anaerobic rumen fungus: Neocallimastix joyonii 总被引:2,自引:0,他引:2
André Breton Annick Bernalier Frédérique Bonnemoy Gérard Fonty Brigitte Gaillard Philippe Gouet 《FEMS microbiology letters》1989,58(2-3):309-314
A new strain of strictly anaerobic fungi was isolated from the rumen of sheep. This strain is characterized by a polycentric thallus, an extensive and polynuclear rhizomycelium, polyflagellated zoospores with gamma particle-like bodies. We propose to assign this strain in a new species: Neocallimastix joyonii. 相似文献
26.
Brigitte Huss Bruno Tinland François Paulus Bernard Walter Léon Otten 《Plant molecular biology》1990,14(2):173-186
The ubiquitous grapevine-associated octopine/cucumopine Ti plasmids of biotype III Agrobacterium tumefaciens strains carry two T regions, TA and TB, with a complex oncogene arrangement. Within the octopine/cucumopine group, two main strain types were identified: large TA strains with a TA region resembling the TL region of the biotype I octopine strain Ach5 and small TA strains with a similar T region organization as the large TA strains but with a large internal TA deletion. Structural and functional studies of the representative large TA strain Tm4 revealed six oncogenes. Each oncogene was inserted in a disarmed vector and tested for biological activity using the corresponding oncogenes of Ach5 as standards. Five Tm4 oncogenes, TA-iaaM, T-ipt, T-6b, TB-iaaH and TB-iaaM, were shown to be active, the IS-interrupted TA-iaaH gene was inactive. To study the role of each gene in the pTiTm4 context, several single and multiple pTiTm4 mutations were constructed. It was shown that whereas TA-iaaM and TB-iaaH are essential for tumour formation on grapevine, T-ipt, T-6b and TB-iaaM are not. The avirulence of the TA-iaaM
-
mutant was shown to be due to an inhibitory effect of the T-ipt gene, since a TA-iaaM
-
/T-ipt
-
double mutant was fully virulent. We conclude that the TA-iaaM gene of large TA strains is specifically required to counteract the tumour growth inhibiting activity of the T-ipt gene. Both TA-iaaM and T-ipt are absent from the small TA strains. A model on the roles and interactions of the different oncogenes in large TA and small TA strains is presented. 相似文献
27.
Patricia Passilly Brigitte Jannin Shawn J. Hassell Norbert Latruffe 《Experimental cell research》1996,223(2):436
Peroxisome proliferators, and especially hypolipidemic drugs such as ciprofibrate, are known to be hepatocarcinogens in rodents, but their effect in humans is controversial. In an attempt to investigate the effects of ciprofibrate at a cellular level, the analysis of individual whole cells was performed by flow cytometry on samples from two hepatic-derived cell lines: the rat Fao cell line and the human HepG2 cell line. The increase of light scatter signals in rat Fao cells treated for 3 days with ciprofibrate at 250 μMwas related to modifications of intrinsic cellular parameters, such as size and cytoplasmic granularity. Conversely, no variations appeared in human HepG2-treated cells. Moreover, the study of the cell cycle distribution of asynchronously growing cells showed an increase in the percentage of proliferative cells in Fao-treated cells, but not in HepG2-treated cells. In order to give a simultaneous assessment of changes in cellular parameters and cell metabolism, these flow cytometric experiments were completed with the measurements of the palmitoyl–CoA oxidase activity, used as a marker of peroxisome proliferation. The cellular modifications in the rat Fao cell line were accompanied by a great increase in this enzymatic activity, whereas the human HepG2 cell line, which failed to exhibit changes of cytometric data, presented no, or weak, increase in this oxidase activity. The cellular modifications observed in the rat Fao cell line may be related to the well-known hepatocarcinogenicity of ciprofibrate in rodents, whereas the absence of response of HepG2 cells is in favor of the noncarcinogenicity of this drug in humans. This report validates another methodological approach for the investigation of the safety of peroxisome proliferators in humans. 相似文献
28.
Hoch Brigitte Lutsch Gudrun Schlegel Wolfgang-Peter Stahl Joachim Wallukat Gerd Bartel Sabine Krause Ernst-Georg Benndorf Rainer Karczewski Peter 《Molecular and cellular biochemistry》1996,160(1):231-239
Recent investigations concentrate on the correlation between the myocardial expression of the inducible 70-kDa heat shock protein (HSP70i) by different stress conditions and its possible protective effects. Only few studies have focused on the involvement of small heat shock proteins in this process. We analyzed the location of the small heat shock protein HSP25 in isolated cardiomyocytes as well as its location and induction in isolated perfused hearts of rats. By immunofluorescence microscopy HSP25 was found to colocalize with actin in the I-band of myofibrils in cardiomyocytes of isolated perfused hearts as well as in isolated neonatal and adult cardiomyocytes. Hyperthermic perfusion of isolated hearts for 45 min resulted in modulation of different parameters of heart function and in induction of HSP25 and HSP70i. Temperatures higher than 43°C (44–46°C) were lethal with respect to the contractile function of the hearts. Compared to control hearts perfused at 37°C, significant increases during hyperthermic perfusion at 42°C and 43°C were obtained for heart rate, contraction velocity and relaxation velocity. In response to hyperthermia at 43°C and after subsequent normothermic perfusion for 135 min at 37°C, left ventricular pressure, contraction velocity and relaxation velocity remained significantly elevated. However, heart rate returned to control values immediately after the period of heat treatment. HSP25 is constitutively expressed even in normothermic perfused hearts as shown by Western blotting. Hyperthermia increased the content of HSP25 only in the left ventricular tissue. In contrast, HSP70i was strongly induced in all analyzed parts of the myocardium (left ventricle, right ventricle, septum). Our findings suggest a differential regulation of HSP25 and HSP70i expression in response to hyperthermia in isolated perfused hearts. The constitutively expressed HSP25 seems to be located adjacent to the myofibrils which implies a specific role of this protein even under unstressed conditions for the contractile function of the myocardium. 相似文献
29.
Yves Paquette Yannick Merlen Brigitte Malette Gilles Bleau 《Molecular reproduction and development》1995,42(4):388-396
Oviductins are high-molecular-weight glycoproteins specifically secreted by the oviduct. These proteins bind to the zona pellucida of the ovulated oocyte and remain associated with the embryo during its transit in the oviduct. They may be involved in fertilization and early embryonic development. In order to explore their putative biological function, the cDNA sequence corresponding to oviductin in the golden hamster was determined. We found that the deduced amino acid sequence of this heavily O-glycosylated protein presents characteristics typical of mucins, including serine- or threonine-rich tandem repeats. Analysis of several cDNA clones and of genomic DNA revealed the presence of a single copy gene with two frequent alleles differing in the number of repeats. Comparison with oviductin sequences from other mammals indicates a high degree of conservation amongst species, except for the repeat region which shows divergence, notably in the number of repeats. Based on its biochemical and genetic properties, hamster oviductin can now be classified as a secretory mucin. This concept provides a new insight in the elucidation of its biological role: oviductin could possibly provide the oviduct and the oocyte with a protective coating ensuring normal tubal function and embryonic development. © 1995 wiley-Liss, Inc. 相似文献