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41.
Barbara Baldan Flavia Guzzo Francesco Filippini Marine Gasparian Fiorella LoSchiavo Alessandro Vitale Sacco C. de Vries Paola Mariani Mario Terzi 《Planta》1997,203(3):381-389
The carrot cell variant ts11 is unable to form somatic embryos at the non-permissive temperature of 32 °C, but the block
can be overcome by the addition of a 32-kDa acidic endochitinase to the medium. In this work we conducted a cyto-histological
analysis of the blocked embryo forms. The morphology of the endomembrane system is altered; in particular, the ER is dilated
and may show electron-dense precipitates and continuity with the plasma membrane. These morphological alterations do not occur
in the presence of externally-added endochitinase. We also noticed modifications of the culture medium that are probably related
to the morphological observations: the total amount of secreted proteins is reduced and pulse-chase experiments revealed that,
compared with wild-type cells, the secretion of major polypeptides is reduced while new minor polypeptides are secreted. Western
blot analysis revealed the presence of the binding protein BiP, a resident of the ER and of glutamine synthase, a cytosolic
protein, in the medium of ts11 but not wild-type cells. These results indicate that ts11 is altered in the secretory pathway
but do not clarify the role of endochitinase.
Received: 13 January 1997 / Accepted: 21 March 1997 相似文献
42.
AV Bezrukov 《Biotechnic & histochemistry》2017,92(1):29-35
I give an historical account and analysis of the scientific priority of the discovery of the polychrome staining of microscopic biological preparations provided by mixtures of eosin plus methylene blue and its derivatives, especially azure B. I maintain that both the formal priority for the discovery of the polychrome staining phenomenon and credit for initiating the development of a technique of polychrome staining properly belong to D. L. Romanowsky. His scientific work demonstrated the possibility of using a simple technique to stain hematological preparations selectively to give good contrast, high resolution and the ability to identify malaria parasites. Romanowsky’s approach constituted the starting point for the development of a family of polychrome stains for microscopic investigation of hematological preparations by a number of his contemporaries. 相似文献
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46.
Gasparian ME Bychkov ML Dolgikh DA Kirpichnikov MP 《Protein expression and purification》2011,79(2):191-196
Enteropeptidase (synonym: enterokinase, EC 3.4.21.9) is a heterodimeric serine protease of the intestinal brush border that activates trypsinogen by highly specific cleavage of the trypsinogen activation peptide following the sequence (Asp)(4)-Lys. It has also great biotechnological interest because of the unique substrate specificity of the serine protease domain. The high degree of specificity exhibited by enteropeptidase makes it a suitable reagent for cleaving recombinant proteins to remove affinity or other tags. However often unwanted cleavages elsewhere in the protein occurred during cleavage of fusions when high amount of enzyme is required. In this study we have improved the efficiency of fusion proteins cleavage by enteropeptidase by substitution of the Lys residue by Arg in specific cleavage sequence (Asp)(4)-Lys. We have demonstrated that 3-6-fold lower amounts of the catalytic subunit of human and bovine enteropeptidase is required for 95% cleavage of Trx/TRAIL and Trx/FGF-2 fusions with (Asp)(4)-Arg cleavage sequence in comparison to native sequence (Asp)(4)-Lys. As a result, reduced amount of non-specifically cleaved peptide fragments were observed during cleavage of (Asp)(4)-Lys/Arg mutated fusions. These findings overcome limitations of enteropeptidase in tag removal processes during recombinant proteins purification and extend its commercial benefit in the biopharmaceutical industry. 相似文献
47.
A flavin-containing polyamine oxidase was isolated in an electrophoretically homogeneous state from cattle liver cytosol. The molecular mass, subunit composition and flavin content of the enzyme were determined; flavin, was shown to be covalently bound to the protein fragment of the polyamine oxidase molecule. Some optical and luminescent properties of the native and denatured enzyme were investigated. Denaturation and quenching were found to affect the luminescent properties of polyamine oxidase. 相似文献
48.
Comparison of amylolytic properties of Lactobacillus amylovorus and of Lactobacillus amylophilus 总被引:2,自引:0,他引:2
Carlos Castillo Pompeyo Maricela Suárez Gómez Sylvie Gasparian Juliette Morlon-Guyot 《Applied microbiology and biotechnology》1993,40(2-3):266-269
Partially purified amylases produced by Lactobacillus amylovorus and L. amylophilus were compared and they differed in several properties. The maximum amylase activity of L. amylovorus was higher than that of L. amylophilus. As estimated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, the molecular mass of the enzymes was 140 kDa for L. amylovorus amylase and 100 kDa for L. amylophilus amylase. Maximum enzymatic activities were obtained when the strains were grown in the presence of CaCO3, on maltose with L. amylovorus and on sucrose with L. amylophilus. Optimal activities were obtained at pH values between 5.0 and 6.0 for both amylases. The L. amylovorus amylase was stable at a higher temperature (50°C) than the L. amylophilus amylase (40°C). Of six substrates examined, greatest activity was obtained by both enzymes on soluble starch. Neither enzymes hydrolysed pullulan or - and \-cyclodextrins. With the exception of Hg2+, which partially inhibited both enzymes, various metal ions, such as 1 mm Ca2+ and Ba2+, stimulated L. amylophilus amylase activity whereas they inhibited L. amylovorus amylase activity.
Correspondence to: J. Morlon-Guyot, ORSTOM 相似文献
49.
In cell suspension of Desulfovibrio desulfuricans B-1388, oxidation of CO as the only energy source is associated with reduction of SO42-. After a 2-h incubation of cells in 8% CO, 81% of the gas is converted. Oxidation of 1 mole CO results in formation of 0.23 mole H2S. Intracellular ATP content increases from 2.5 (control) to 8.3 nmoles/mg (during CO conversion). Dinitrophenol inhibits sulfate reduction and CO oxidation. CO dehydrogenase was detected in cytoplasmic and membrane cell fractions (59 and 34%, respectively). 相似文献
50.
Gasparian ME Domnina LV Ivanova OY Izyumov DS Lomakin AY Popova EN Yagolovich AV Pletjushkina OY Dolgikh DA Chernyak BV 《Biochemistry. Biokhimii?a》2008,73(3):358-362
TRAIL (Apo2L), a cytokine from the family of tumor necrosis factors (TNF), causes apoptosis in various types of tumor cells but is not toxic for normal cells. Recombinant TRAIL obtained using an original method stimulates the release of cytochrome c from mitochondria into the cytoplasm and apoptosis in HeLa carcinoma cells. Expression of oncoprotein Bcl-2 in these cells blocks both processes. The microtubule inhibitors taxol, nocodazole, and colcemid, as well as an inhibitor of actin microfilaments cytochalasin D, enhance the action of TRAIL and allow it to overcome protection caused by overexpression of Bcl-2. This effect is not associated with enhancement of early steps of TRAIL-dependent apoptosis leading to activation of caspase-8 and Bid protein. The inactivation of Bcl-2 also does not define the effect of cytoskeleton inhibitors. It is supposed that destruction of cytoskeleton alters the mechanism of the TRAIL- (or TNF)-dependent cytochrome c release from mitochondria by making it resistant to Bcl-2. The combined use of cytoskeleton inhibitors, which are antitumor drugs, with the recombinant TRAIL preparations may be efficient in therapy of tumors resistant to traditional chemotherapy. 相似文献