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101.
H. Yanase M. Okuda K. Kita Y. Sato K. Shibata Y. Sakai N. Kato 《Applied microbiology and biotechnology》1995,43(2):228-234
Dihydroxyacetoone synthase (EC 2.2.1.3), which is a key enzyme of the C1-compound-assimilating pathway in yeasts, catalyzes transketolation between formaldehyde and hydroxypyruvate, leading to the formation of dihydroxyacetone and CO2. When [13C]formaldehyde was used as a substrate with dihydroxyacytone synthase from Candida boidinii 2201, 13C was confirmed to be incorporated to the C-1 and C-3 positions of dihydroxyacetone, and the 13C content of each carbon (atoms/100 atoms) was estimated to be 50%. [13C]Methanol was also useful for the enrichment of dihydroxyacetone with 13C, when alcohol oxidase from a methylotrophic yeast was added for the conversion of methanol to formaldehyde. A fed-batch reaction with periodic addition of the substrates was required for the accumalation of 13C-labelled dihydroxyacetone at a higher concentration, because the enzyme system was relatively susceptible to the C donor, formaldehyde or methanol. The optimum conditions for the production gave 160mM (14.4 mg/ml) dihydroxyacetone for 180 min; the molar yield relative to methanol added was 80%. Diyhdroxyacetone kinase (EC 2.7.1.29) from methanol-grown Hansenula polymorpha CBS 4732 was a suitable enzyme for the phosphorylation of dihydroxyacytone. The phosphorylation system, comprising of dihydroxyacetone kinase, adenylate kinase, and ATP, could be coupled with the system for dihydroxyacetone production. A fed-batch reaction afforded 185 mM [1, 3-13C]dihydroxyacetone phosphate from [13C]methanol; the molar yield of the ester relative to methanol added was 92.5% 相似文献
102.
Electrophysiological properties of mouse bone marrow-derived mast cells (BMMC) were studied under the whole-cell clamp configuration. About one third of the cells were quiescent, but others expressed either inward or outward currents. Inwardly rectifying (IR) currents were predominant in 14% of the cells, and outwardly rectifying (OR) currents in 24%. The rest (22%) of the cells exhibited both inward and outward currents. The IR currents were eliminated by 1 mm Ba2+, and were partially inhibited by 100 μm quinidine. The reversal potential was dependent on extracellular K+, thereby indicating that K+ mediated the IR currents. The negative conductance region was seen at potentials positive to E K. The OR currents did not apparently depend on the extracellular K+ concentration, but were reduced by lowering the extracellular Cl? concentration. The OR currents were partially blocked by 1 mm Ba2+, and were further blocked by a Cl? channel blocker, 4,4′-diisothiocyano-2, 2′-stilbenedisulfonate (DIDS). In addition, the reversal potential of the OR currents was positively shifted by decreasing the ratio of external and internal Cl? concentrations, suggesting that Cl? was a major ion carrier. In cells exhibiting IR currents, the membrane potential varied among cells and tended to depolarize by elevating the external K+ concentration. In cells with OR currents, the resting potential was hyperpolarized in association with an increase in conductance. These results suggest that BMMC have a heterogeneous electrophysiological profile that may underlie a variety of ion channels expressed in different phenotypes of mast cells. Activities of both the inwardly rectifying K+ channel and the outwardly rectifying Cl? channel seem to contribute to the regulation of the membrane potential. 相似文献
103.
Seitaro Ohkuma Hidehiko Narihara Masashi Katsura Takeshi Hasegawa Kinya Kuriyama 《Journal of neurochemistry》1995,65(3):1109-1114
Abstract: The functional significance of peroxynitrite in the release of [3 H]GABA induced by nitric oxide (NO) liberated from NO generators was investigated using cerebral cortical neurons in primary culture. NO generators such as sodium nitroprusside (SNP) and S -nitroso- N -acetylpenicillamine (SNAP) increased [3 H]GABA release in a dose-dependent manner. These increases in [3 H]GABA release were significantly inhibited by hemoglobin, indicating that those NO generators evoke the release of [3 H]GABA by the formation of NO. Two types of superoxide scavengers, Cu2+ /Zn2+ superoxide dismutase and ceruloplasmin, significantly reduced the increase in [3 H]GABA release induced by both SNP and SNAP, which assumes that NO requires superoxide to induce [3 H]GABA release from the neurons. In addition, synthesized peroxynitrite induced a dose-dependent increase in [3 H]GABA release from the neurons. These results indicate that NO-induced [3 H]GABA release is mediated by peroxynitrite formed by the reaction of NO with superoxide. 相似文献
104.
Changes in chorion proteins induced by the exudate released from the egg cortex at the time of fertilization in the teleost, Oryzias latipes 总被引:2,自引:2,他引:0
Takashi Iwamatsu Yasushi Shibata Toshihiro Kanie 《Development, growth & differentiation》1995,37(6):747-759
The chorion of unfertilized medaka Oryzias latipes eggs consists of two major proteins (77–73 and 49 kDa) and a minor 150 kDa protein. Upon fertilization, these major chorion proteins are polymerized to insoluble high molecular weight proteins via the temporary formation of several new proteins (132, 114, 62 and 61 kDa). Increasing chorion toughness is closely related to the formation of high molecular weight proteins and the increasing insolubility of the chorion proteins. The changes in chorion proteins and hardening could be induced in vitro in isolated chorions by an egg exudate, which includes cortical alveolar contents. The effects of temperature and pH on the egg exudate-induced changes in chorion proteins were examined in the present study. The major proteins could be digested by proteolytic enzymes. The 49 kDa protein was PAS-positive. Analysis with polyclonal antibodies against the major proteins demonstrated that the temporarily formed 62 and 61 kDa proteins were derived from the 77–73 kDa protein and that higher molecular weight proteins, newly formed in the process of chorion hardening, contained the same epitopes as did the 77–73 and 49 kDa proteins. The results suggest that the changes in chorion proteins of the medaka egg at the time of fertilization can be induced by an enzyme(s) released from the egg cortex into the perivitelline space. 相似文献
105.
Molecular Evolution of the Duplicated Amy Locus in the Drosophila Melanogaster Species Subgroup: Concerted Evolution Only in the Coding Region and an Excess of Nonsynonymous Substitutions in Speciation
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From the analysis of restriction maps of the Amy region in eight sibling species belonging to the Drosophila melanogaster species subgroup, we herein show that the patterns of duplication of the Amy gene are almost the same in all species. This indicates that duplication occurred before speciation within this species subgroup. From the nucleotide sequence data, we show a strong within-species similarity between the duplicated loci in the Amy coding region. This is in contrast to a strong similarity in the 5' and 3' flanking regions within each locus (proximal or distal) throughout the species subgroup. This means that concerted evolution occurred only in the Amy coding region and that differentiated evolution between the duplication occurred in the flanking regions. Moreover, when comparing the species, we also found a significant excess of nonsynonymous substitutions. In particular, all the fixed substitutions specific to D. erecta were found to be nonsynonymous. We thus conclude that adaptive protein evolution occurred in the lineage of D. erecta that is a ``specialist' species for host plants and probably also occurs in the process of speciation in general. 相似文献
106.
Mercurial-sensitive water transport in barley roots 总被引:16,自引:0,他引:16
Masashi Tazawa Eiji Ohkuma Mineo Shibasaka Susumu Nakashima 《Journal of plant research》1997,110(4):435-442
An isolated barley root was partitioned into the apical and basal part across the partition wall of the double-chamber osmometer.
Transroot water movement was induced by subjecting the apical part to a sorbitol solution, while the basal part with the cut
end was in artificial pond water. The rate of transroot osmosis was first low but enhanced by two means, infilitration of
roots by pressurization and repetition of osmosis. Both effects acted additively. The radial hydraulic conductivity (Lpr) was calculated by dividing the initial flow rate with the surface area of the apical part of the root, to which sorbitol
was applied, and the osmotic gradient between the apical and basal part of the root. Lpr which was first 0.02–0.04 pm s−1 Pa−1 increased up to 0.25–0.4 pm s−1 Pa−1 after enhancement. Enhancement is assumed to be caused by an increase of the area of the plasma membrane which is avallable
to osmotic water movement. The increased Lpr is in the same order of magnitude as the hydraulic conductivity (Lp) of epidermal and cortical cells of barley roots obtained
by Steudie and Jeschke (1983). HgCl2, a potent inhibitor of water channels, suppressed Lpr of non-infiltrated and infiltrated roots down to 17% and 8% of control values, respectively. A high sensitivity of Lpr to HgCl2 suggests that water channels constitute the most conductive pathway for osmotic radial water movement in barley roots. 相似文献
107.
Alexander A. Bachmanov Danielle R. Reed Yuro Ninomiya Masashi Inoue Michael G. Tordoff R. Arlen Price Gary K. Beauchamp 《Mammalian genome》1997,8(8):545-548
Individual variability in sucrose consumption is prominent in humans and other species. To investigate the genetic contribution
to this complex behavior, we conducted behavioral, electrophysiological, and genetic studies, using male progeny of two inbred
mouse strains (C57BL/6ByJ [B6] and 129/J [129]) and their F2 hybrids. Two loci on Chromosome (Chr) 4 were responsible for over 50% of the genetic variability in sucrose intake. These
loci apparently modulated intake by altering peripheral neural responses to sucrose. One locus affected the response threshold,
whereas the other affected the response magnitude. These findings suggest that the majority of difference in sucrose intake
between male B6 and 129 mice is due to polymorphisms of two genes that influence receptor or peripheral nervous system activity.
Received: 27 January 1997 / Accepted: 17 March 1997 相似文献
108.
Isolation and characterization of Enterobacter cloacae mutants which are defective in chemotaxis toward inorganic phosphate. 总被引:1,自引:0,他引:1
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Enterobacter cloacae IFO3320 is attracted to Pi when cells are starved for Pi. Two Tn1737KH-induced mutants, which were constitutive for alkaline phosphatase, failed to exhibit Pi taxis even under conditions of Pi limitation. Both of the mutant strains exhibited normal chemotactic responses to peptone, suggesting that they are specifically defective in Pi taxis. Cloning and sequence analysis showed that the TN1737KH insertions were located in either the pstA or pstB genes which encode the channel-forming proteins of the Pi-specific transport (Pst) system in E. cloacae. These results suggest that the E. cloacae Pst system is required for Pi chemoreception. 相似文献
109.
The relationship between adenine-nucleotide levels and metabolism-dependent membrane potential was studied in cells of Nitellopsis obtusa. Effects of ADP and AMP in the presence of ATP on electrogenic pump activity were measured in the dark, using the continuous perfusion method. Both ADP and AMP acte as competitive inhibitors for ATP, the Ki value for either compound being about 0.4 mM. The role of ADP and AMP as regulating factors for the electrogenic pump was investigated under various metabolic conditions. Application of N2 gas in the dark caused a significant membrane depolarization amounting to 90 mV, but cytoplasmic streaming and membrane excitability were not affected. Under anoxia, the ATP level decreased from 1.6 to 0.5 mM; ADP increased but only slightly, and AMP increased greatly. However, the time course of changes in the adenine nucleotides was not concurrent with that of the membrane-potential changes, thus, the adenine-nucleotide level changes cannot fully account for the N2-elicited depolarization. Under light, although the membrane hyperpolarized, no significant changes in the adenine-nucleotide levels were observed. Therefore, the light-induced membrane hyperpolarization cannot be explained solely by changes in adenine-nucleotide levels.Abbreviations APW
artificial pond water
- Em
membrane potential
- Rm
membrane resistance 相似文献
110.
The substrate specificity of rice embryo benzoyl-L-argininep-nitroanilide hydrolase (BAPAase) was examined. No endopeptidaseactivity toward protein substrates was detectable. Small peptides(less than 8 residues) and amide, ester substrates, however,were hydrolyzed very well at the carboxyl side of the lysineor arginine residue. No other peptide bond was hydrolyzed. TheN-terminal arginine of the substrates was released very slowly.Peptides with lysine or arginine penultimate to the C-terminalposition were hydrolyzed well and released an amino acid. Theoxidized insulin B chain (30 residues) was cleaved very slowlyat the C-terminal Lys-Ala bond, whereas an Arg-Gly bond at aninner position was not cleaved. The hydrolytic rate increasedafter the chain length was shortened by chymotryptic digestion.These results show that the rice embryo BAPAase is a novel enzymewhich has mixed endopeptidase-carboxypeptidase activity towardthe Arg-X and Lys-X bonds of small peptides, a characteristicintermediate between trypsin and serine carboxypeptidase. Thisenzyme may act in the breakdown of small peptides that havephysiological functions. (Received May 26, 1984; Accepted August 29, 1984) 相似文献