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1.
The KdpFABC complex of Escherichia coli, which belongs to the P-type ATPase family, has a unique structure, since catalytic activity (KdpB) and the capacity to transport potassium ions (KdpA) are located on different subunits. We found that fluorescein 5-isothiocyanate (FITC) inhibits ATPase activity, probably by covalently modifying lysine 395 in KdpB. In addition, we observed that the KdpFABC complex is able to hydrolyze p-nitrophenyl phosphate (pNPP) in a Mg(2+)-dependent reaction. The pNPPase activity is inhibited by FITC and o-vanadate. Low concentrations of ATP (1-30 microM) stimulate the pNPPase activity, while concentrations of >500 microM are inhibitory. This behavior can be explained either by a regulatory ATP binding site, where ATP hydrolysis is required, or by proposing an interactive dimer. The notion that FITC inhibits pNPPase and ATPase activity supports the idea that the catalytic domain of KdpB is much more compact than other P-type ATPases, like Na(+),K(+)-ATPase, H(+),K(+)-ATPase, and Ca(2+)-ATPase. 相似文献
2.
Summary The influence of O2-deprivation and reduction of temperature on the incorporation of the RNA-precursors 3H-uridine und 3H-cytidine is investigated in various tissues of the larvae and in the ovaries of adults of the housefly Musca domestica L. While RNA-synthesis in most of the tissues is strongly reduced under anaerobic conditions, synthesis continues in a moderate extent in muscle cell nuclei and nurse cell nuclei. The RNA-macromolecules (mRNA and rRNA), however, do not migrate into the cytoplasma. RNA-synthesis within the cell nucleus is less affected by a sudden reduction of temperature than the passage of RNA through the nuclear membrane which is reduced to a very low rate. The macromolecular RNA, therefore, does not diffuse into the cytoplasma but is transported actively through the nuclear envelope. The malformations caused by anaerobiosis during embryogenesis are brought in connexion with the active RNA-transport through the nuclear envelope and the separation of transport and synthesis.
Herrn Professor Dr. Hans Bauer zum 60. Geburtstag gewidmet.
Mit Unterstützung durch die Deutsche Forschungsgemeinschaft. 相似文献
Herrn Professor Dr. Hans Bauer zum 60. Geburtstag gewidmet.
Mit Unterstützung durch die Deutsche Forschungsgemeinschaft. 相似文献
3.
Karlheinz Mann Jo?l Gautron Yves Nys Marc D McKee Tarek Bajari Wolfgang J Schneider Maxwell T Hincke 《Matrix biology》2003,22(5):397-407
Clusterin is a widely expressed secretory glycoprotein which is found in mammals as a disulfide-bonded alpha/beta heterodimer generated by cleavage of the single-chain precursor. In contrast, clusterin occurs in the chicken mainly as an intracellular single-chain form and is not observed in serum. The present report identifies chicken clusterin as a component of the eggshell. This extracellular clusterin originates in the uterine fluid, where it is a disulfide-bonded heterodimer derived from the precursor polypeptide by proteolytic cleavage at the same site as in mammals. Clusterin message expression in the oviduct was measured by real time RT-PCR, and levels were found to be highest in magnum and uterus. Western blotting using protein extracts of oviduct tissues indicated major clusterin production in the magnum, while immunostaining of the oviduct identified clusterin in the tubular glands of the uterus and the magnum. In addition, clusterin was detected in egg white by Western blotting. In the decalcified eggshell, immunofluorescence and colloidal-gold immunocytochemistry revealed that clusterin was predominantly localized in the palisade and mammillary layers, but also in the mantle and core of the inner and outer shell membranes. It has been suggested recently that clusterin acts as an extracellular chaperone. Thus clusterin could function in the uterine fluid to prevent the premature aggregation and precipitation of eggshell matrix components before and during their assembly into the rigid protein scaffold necessary for ordered mineralization. 相似文献
4.
Saccharomyces cerevisiae nuclear membranes were prepared from isolated nuclei by digesting chromatin with deoxyribonuclease and ribonuclease, washing of residual nuclei with 0.5 M MgCl2, and discontinuous gradient centrifugation in buffered Ficoll solutions. Electron microscopic examination of the preparations showed single membrane and double membrane vesicles and membrane sheets. Pores or residual pores were often visible. In double membrane profiles the two unit membranes were often separated by the remains of the perinuclear cistern. The nuclear membrane fragments contained 58% protein, 23.8% phospholipid, 6% sterols, 7.1% neutral acylglycerols, 4.8% RNA, and 0.3% DNA. The phospholipid content of the membrane preparations was influenced by a phospholipase activity with acidic pH optimum. 相似文献
5.
Takako Sasaki Karlheinz Mann Jeffrey H Miner Nicolai Miosge Rupert Timpl 《European journal of biochemistry》2002,269(2):431-442
Domain IV, consisting of about 230 residues, represents a particular protein module so far found only in laminin beta1 and beta2 chains. Both domains were obtained by recombinant production in mammalian cells. They showed a globular structure, as expected from electron microscopic examination of laminins. Fragment beta1IV was obtained as a monomer and a disulfide-bonded dimer, and both were modified to approximately 50% by a single chondroitin sulfate chain attached to Ser721 of an SGD consensus sequence. Dimerization is caused by an odd number of cysteines, with three of them having a partial thiol character. Whether both modifications also occur in tissue forms of laminin remains to be established. Fragment beta2IV was only obtained as a monomer, as it lacked one crucial cysteine and the SGD sequence. It required, however, the presence of two adjacent LE modules for proper folding. Polyclonal antibodies raised against both fragments showed no cross-reaction with each other and allowed establishment of beta chain-specific radioimmunoassays and light and electron microscopic immunostaining of tissues. This demonstrated a 5-25-fold lower content of beta2 compared with beta1 chains in various tissue extracts of adult mice. Tissues derived from beta2-deficient mice failed to react with the beta2-specific antibodies but showed a twofold higher content of beta1 than heterozygotes. The antibodies to beta2 showed broader tissue staining than reported previously, including in particular a distinct reaction with the extrasynaptic endomysium of skeletal muscle. Immunogold staining localized both beta chains primarily to basement membranes of kidney, muscle and various other tissues. 相似文献
6.
B Schneppe G Deckers-Hebestreit K Altendorf 《The Journal of biological chemistry》1990,265(1):389-395
The uncI gene, the first gene of the unc operon, has been cloned into an expression vector carrying the lambda PRPL promoters in tandem orientation and the gene cI857 coding for the thermolabile repressor. Linkage of the uncI gene to an efficient ribosome binding site (the translational initiation region of the uncE gene) resulted in 10-20-fold increased gene expression. The i protein has been extracted from overproducing cells using chloroform/methanol and purified to homogeneity by ion exchange chromatography. Analyzing the products of the uncI gene encoded by different plasmids, we provide evidence that, in contrast to the previously reported data (Walker, J. E., Saraste, M., and Gay, N. J. (1984) Biochim. Biophys. Acta 768, 164-200), the chromosome-encoded i protein contains the N-terminal sequence Ser-Val-Ser-Leu-Val-Ser-Arg and has a molecular weight of 13,504. 相似文献
7.
Zusammenfassung Mit Hilfe von autoradiographischen und elektrophoretischen Methoden wurde die Dottereinlagerung in den wachsenden Oocyten vonMusca domestica untersucht. Sie beginnt nach 30 min im Autoradiogramm sichtbar zu werden. Durch ihre Färbbarkeit und Markierung konnte die Dotterfraktion im Pherogramm von Ovar und Hämolymphe eines mittleren Wachstumsstadiums (Stadium 3) nachgewiesen werden. Nach Abschlu der Vitellogenese tritt sie in der Hämolymphe nicht mehr auf. Die Einlagerung der Dotterproteine wird durch Actinomycin gestört, dagegen läuft ihre Synthese nahezu unbeeinflut weiter. Die Transporthemmung kann als bisher unbekannter Nebeneffekt des Actinomycins gedeutet werden.
Synthesis of haemolymph proteine and the uptake of the yolk fraction in the oocyte during Actinomycin-treatment. (Studies onMusca domestica)
Summary By means of radioautographic and electrophoretic techniques yolk protein uptake in the growing oocytes ofMusca domestica was investigated. After 30 min yolk protein becomes visible in the radioautograms. By stainability and labeling the yolk fraction could be demonstrated in the pherogram of ovary and haemolymph in an intermediate developmental stage (stage 3). After the end of vitellogenesis it does not appear in the haemolymph. The yolk protein uptake is inhibited by Actinomycin, but the synthesis goes on nearly as normal. This inhibition can be interpretated as a new accessory effect of Actinomycin.相似文献
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9.
Patil Satish Sawant Sarvesh Hauff Karlheinz Hampp Gabriele 《Probiotics and antimicrobial proteins》2019,11(4):1124-1131
Probiotics and Antimicrobial Proteins - Hylak® forte is a postbiotic that inhibits the growth of pathogenic bacteria by reducing intestinal pH. It is assumed the potential presence of... 相似文献
10.