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951.
Cytosolic protein phosphotyrosine phosphatases from rabbit kidney. Purification of two distinct enzymes that bind to Zn2+-iminodiacetate agarose 总被引:14,自引:0,他引:14
Cytosolic protein phosphotyrosine (PPT) phosphatase was measured using a new substrate, Tyr(32P)-labeled bovine serum albumin. Kidney was found as a particularly rich tissue source of PPT-phosphatase activity, containing twice as much as liver and over 10-fold more than brain, heart, lung, or skeletal muscle. An affinity column of Zn2+-iminodiacetate agarose adsorbed up to 60% of the PPT-phosphatase present in kidney extracts. Subsequent chromatography on DEAE-Sepharose separated the phosphatase into two peaks, labeled I and II, that had Mr = 34,000 and 37,000, respectively, upon gel filtration with Sephadex G-75 Superfine. Overall purification of 850- and 1100-fold was achieved with a net 4% yield. Both phosphatases hydrolyzed p-nitrophenylphosphate as well as the protein substrate in the presence of EDTA. Peak I phosphatase activity displayed a neutral pH optimum, had an absolute requirement for sulfhydryl compounds, and was sensitive to trypsin, whereas Peak II activity had an acidic pH optimum and was active without mercaptans. The two proteins also gave different fragmentation patterns by gel electrophoresis after digestion with S. aureus V8 protease. The results show that multiple forms of PPT phosphatase specifically interact with Zn2+ and provide a basis for further structural and functional comparisons among different members of the phosphoprotein phosphatase family. 相似文献
952.
953.
954.
F Kh Sharipov 《Biulleten' eksperimental'no? biologii i meditsiny》1985,100(8):245-247
It has been established in experiments on 280 white randombred male rats weighing 100-120 g that the lifting of the animals from the valley (820 m above the sea level) to the mountains (3379 m above the sea level) brings about within the first day marked suppression of the mitotic activity of the epithelium of involuted renal tubules. This activity increases beginning from the end of the first week, approaches the control value by the 30th day of adaptation and almost completely returns to normal by the 60th day of the animals' stay in the mountains. The circadian rhythms of the mitotic activity appeared undisturbed and was monophasic in nature. 相似文献
955.
The basis for the apparent cooperativity in the activation of protein kinase C by phosphatidylserine has been addressed using proteolytic sensitivity, resonance energy transfer, and enzymatic activity. We show that binding of protein kinase C to detergent-lipid mixed micelles and model membranes is cooperatively regulated by phosphatidylserine. The sigmoidal dependence on phosphatidylserine for binding is indistinguishable from that observed for the activation of the kinase by this lipid [Newton & Koshland (1989) J. Biol. Chem. 264, 14909-14915]. Thus, protein kinase C activity is linearly related to the amount of phosphatidylserine bound. Furthermore, under conditions where protein kinase C is bound to micelles at all lipid concentrations, activation of the enzyme continues to display a sigmoidal dependence on the phosphatidylserine content of the micelle. This indicates that the apparent cooperativity in binding does not arise because protein kinase C senses a higher concentration of phosphatidylserine once recruited to the micelle. Our results reveal that the affinity of protein kinase C for phosphatidylserine increases as more of this lipid binds, supporting the hypothesis that a domain of phosphatidylserine is cooperatively sequestered around the enzyme. 相似文献
956.
P N Tonin R L Stallings M D Carman J R Bertino J A Wright P R Srinivasan W H Lewis 《Cytogenetics and cell genetics》1987,45(2):102-108
We have shown previously that cDNAs for the M1 and M2 subunits of ribonucleotide reductase, ornithine decarboxylase (ODC), and p5-8, a 55,000-Dalton protein, hybridize to amplified genomic sequences in a highly hydroxyurea-resistant hamster cell line. We have extended these observations to include two additional, independently isolated, hydroxyurea-resistant cell lines: SC8, a single-step hamster ovary cell line, and KH450, a multistep human myeloid leukemic cell line, have also undergone genomic amplification for sequences homologous to ODC and p5-8 cDNAs. However, neither SC8 nor KH450 contains amplified genomic sequences homologous to an M1 cDNA probe. A panel of mouse-hamster somatic cell hybrids was used to map sequences homologous to M1, M2, ODC, and 5-8 cDNAs in the hamster genome. The M2, ODC, and p5-8 cDNAs hybridized to DNA fragments that segregated with hamster chromosome 7. In contrast, M1 cDNA hybridized to DNA fragments that segregated with hamster chromosome 3. These data suggest that the genes RRM2, (M2), ODC, and p5-8, but not RRMI (M1), are linked and may have been co-amplified in the selection of the hydroxyurea-resistant hamster and human cell lines. 相似文献
957.
958.
Stereospecific binding of 3H-dopamine in neostriatal membrane preparations: inhibitory effects of sodium ascorbate 总被引:2,自引:0,他引:2
It has been pointed out by several different groups of investigators in the past several years that ascorbic acid was a potent inhibitor of the binding of dopamine (DA) agonists including 3H-DA itself and 3H-ADTN, 3H-apomorphine and 3H-norpropylapomorphine to neostriatal membrane preparations. However, the significance of this effect of ascorbic acid has been controversial. For example, it has recently been claimed that the stereospecific binding of DA agonists is facilitated by ascorbic acid and can be measured only in its presence. In the present study in neostriatal membrane preparations in the absence of ascorbic acid, the binding of 3H-DA was very potently inhibited by potent DA agonists (DA, ADTN, apomorphine). Considerably weaker effects were obtained with norepinephrine, isoproterenol, serotonin, catechol and pyrogallol. Stereospecific effects were clearly observed in that the binding of 3H-DA was inhibited to a much greater extent by several biologically active enantiomers than by their less active counterparts. For example, (-)-2-hydroxyapomorphine and (-)-norpropylapomorphine were much more potent inhibitors than their corresponding (+) isomers. This binding of 3H-DA was also very strongly inhibited by sodium ascorbate and several other reducing agents. In control experiments in the neostriatal membrane preparation in the absence of ascorbic acid, there was no detectable decomposition of 3H-DA. The data suggest that 3H-DA can, in the absence of sodium ascorbate, bind stereospecifically to a site that has the properties of a DA receptor. Furthermore, sodium ascorbate is a potent inhibitor of this stereospecific binding. 相似文献
959.
The enzymes of phospholipid synthesis in Clostridium butyricum 总被引:5,自引:0,他引:5
We have examined extracts of Clostridium butyricum for several enzymes of phospholipid synthesis. Membrane particles were shown to catalyze the formation of CDP-diglyceride from [3H]CTP and phosphatidic acid. The reaction was dependent on Mg2+ and stimulated by monovalent cations. CDP-diglyceride formed in vitro was found to be a substrate for both phosphatidylglycerophosphate synthetase and phosphatidylserine synthetase. The formation of phosphatidylglycerophosphate from added CDP-diglyceride and [U-14C]sn-glycerol-3-phosphate was dependent on Mg2+ and Triton X-100. The dephosphorylation of endogenously-generated phosphatidylglycerophosphate to yield phosphatidylglycerol was observed to be pH-dependent. The formation of phosphatidylserine from CDP-diglyceride and L-[3-14C]serine was stimulated by Mg2+ and Triton X-100. dCDP-diglyceride was a suitable substrate for both phosphatidylglycerophosphate synthetase and phosphatidylserine synthetase. Phosphatidylserine decarboxylase activity was barely detectable in membrane particles from C. butyricum. The addition of E. coli membrane particles provided efficient phosphatidylserine decarboxylase activity in this system. Although plasmalogens are the principal lipids of C. butyricum, none of the products of phospholipid synthesis formed in vitro contained measurable amounts of plasmalogens. The subcellular distribution of both phosphatidylglycerophosphate synthetase and phosphatidylserine synthetase in C. butyricum was also studied. Both were found to be membrane-associated. 相似文献
960.
The role of arctic zooplankton in biogeochemical cycles: respiration and excretion of ammonia and phosphate during summer 总被引:1,自引:0,他引:1
M. Alcaraz R. Almeda A. Calbet E. Saiz C. M. Duarte S. Lasternas S. Agustí R. Santiago J. Movilla A. Alonso 《Polar Biology》2010,33(12):1719-1731
The study of the structural and functional properties of key components of polar marine ecosystems has received increased
attention in order to better understand the ecological consequences of future sea temperature rise and seasonal ice retraction.
Owing to this purpose, during the ATOS-Arctic cruise, held in July 2007 in the framework of the 2007–2008 International Polar
Year, we studied the respiratory carbon demand of mesozooplankton as well as their contribution to the regeneration of inorganic
nitrogen and phosphorus (NH4-N and PO4-P) via excretion. The studied area comprised several stations along a latitudinal gradient in the East Greenland current, plus a
network of stations NW of the Svalbard islands. The specific respiratory carbon losses and phosphorus (PO4-P) excretion rates were similar or slightly higher than some reports for Arctic mesozooplankton, but the nitrogen (NH4-N) excretion rates were higher by a factor of 3 when compared with previous data sets. The mesozooplankton respiratory losses
were equivalent to 23% of primary production, and at turn zooplankton contributed by excretion to more than 50% of the N and
P required by phytoplankton. Although C:N, C:P and N:P metabolic atomic quotients almost coincided with the average Redfield’s
stoichiometric ratios, the low C:N values when compared to previous reports suggested a predominance of protein-related metabolic
substrates. The potential consequences of changes observed in the C:N, N:P and C:P metabolic ratios of mesozooplankton for
Arctic marine ecosystems are discussed. 相似文献