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1.
A respiration-deficient (RD) mutant was isolated from the petite-negative, salt-tolerant yeast Zygosaccharomyces rouxii. One strain among sixteen glycerol-non-utilizing mutants exhibited vigorous liberation of CO2 but no uptake of O2. Furthermore, this strain lacked cytochrome aa3 and had a reduced level of cytochrome b. The few mitochondria found in cells of this strain contained few or no cristae. Salt tolerance and intracellular accumulation of glycerol by the RD strain were almost equal to that of the wild-type strain in media containing NaCl up to 2.5 M. In media with more than 3 M NaCl, the growth of the RD mutant was retarded and the intracellular accumulation of glycerol was depressed in spite of ample production. 相似文献
2.
Mutagenesis of human granulocyte colony stimulating factor 总被引:9,自引:0,他引:9
T Kuga Y Komatsu M Yamasaki S Sekine H Miyaji T Nishi M Sato Y Yokoo M Asano M Okabe 《Biochemical and biophysical research communications》1989,159(1):103-111
To define the structure-function relationship, we have made a number of mutants of human granulocyte colony-stimulating factor (hG-CSF) by in vitro mutagenesis. The results indicate that most of the mutations located in the internal and C-terminal regions of the molecule abolished the activity, whereas the mutants without N-terminal 4, 5, 7, or 11 amino acids retained the activity. N-terminal amino acids were also altered by cassette mutagenesis using a synthetic oligonucleotide mixture. Among them, KW2228, in which Thr-1, Leu-3, Gly-4, Pro-5 and Cys-17 were respectively substituted with Ala, Thr, Tyr, Arg and Ser, showed more potent granulopoietic activity than that of intact hG-CSF both in vitro and in vivo. 相似文献
3.
Role of Chitinase and Chitin Oligosaccharides in Lignification Response of Cultured Carrot Cells Treated with Mycelial Walls 总被引:3,自引:0,他引:3
Chitinase activity was induced in cultured carrot cells by incubationwith mycelial walls of a fungus, Chaetomium globosum. Both intra-and extracellular chitinases were resolved into four componentsby gel filtration chromatography. The extracellular enzymesliberated soluble oligosaccharides of different sizes from insolublechitin, suggesting that these carrot chitinases are endo-hydrolases.The solubilized chitinase digests obtained from insoluble mycelialwalls of C. globosum and chitin were fractionated by gel filtrationchromatography, and the elicitor activity of each fraction forthe accumulation of phenolic acids in cultured carrot cellswas determined. In both solubilized fragments of fungal wallsand of chitin, elicitor-active oligosaccharides were distributedin many fractions, however, potent activity for inducing phenolicacid synthesis was observed in the high molecular weight fractions. (Received October 5, 1987; Accepted February 12, 1988) 相似文献
4.
Matuo Y Nishi N Muguruma Y Yoshitake Y Masuda Y Nishikawa K Wada F 《Cytotechnology》1988,1(4):309-318
Among several detergents, a zwitterionic detergent, 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS), was found to be least cytotoxic for cultured mammalian cells. CHAPS improved the activity recovery and elution profile of crude and purified fibroblast growth factors (FGFs) during chromatographies. Diluted preparations of FGFs were stabilized by CHAPS against the loss during storage. Amino acid sequence analysis was not disturbed by CHAPS. CHAPS was removable by reversed-phase high-performance liquid chromatography. These results indicate that CHAPS is useful as a non-cytotoxic stabilizing agent in purification of various kinds of bioactive polypeptides.Abbreviations -MEM
Alpha Modification of Eagle's Minimal essential medium
- CHAPS
3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate
- CHAPSO
3-[(3-cholamidopropyl)dimethylammonio]-2-hydroxy-1-propane sulfonate
- CS
Calf Serum
- EGF
Epidermal Growth Factor
- FGF
Fibroblast Growth Factor
- HPLC
High-Performance Liquid Chromatography
- NGF
Nerve Growth Factor
- NOG
1-O-n-octyl--D-glucopyranoside
- NP-40
Nonidet P-40
- PBS
Phosphate-Buffered Saline
- SB 12
3-(dodecylmethylammonio)-1-propane sulfonate
- SDS
Sodium Dodecyl Sulfate
- TGF- and
Transforming Growth Factor type and 相似文献
5.
Preventive effect of MCI-186 on 15-HPETE induced vascular endothelial cell injury in vitro 总被引:8,自引:0,他引:8
T Watanabe I Morita H Nishi S Murota 《Prostaglandins, leukotrienes, and essential fatty acids》1988,33(1):81-87
Using cultured bovine aortic endothelial cells, the effects of MCI-186, a radical scavenger, were studied on arachidonic acid metabolism and on the cell injury caused by 15-HPETE. MCI-186 at 3 X 10(-5) M enhanced prostacyclin production in the intact endothelial cells without affecting phospholipase A2. When endothelial cell homogenates were used as an enzyme source, it was found that MCI-186 stimulated the conversion of arachidonic acid to prostacyclin like phenol, perhaps by trapping OH radicals produced in the process of the conversion of PGG2 to PGH2. On the other hand, MCI-186 was found to inhibit lipoxygenase metabolism of arachidonic acid in cell free homogenates of rat basophilic leukemia cells. The lipoxygenase inhibition caused by 3 X 10(-5) M MCI-186 was almost equivalent to that caused by 3 X 10(-6) M BW 755C. MCI-186 remarkably protected against endothelial cell damage caused by 15-HPETE. 3 X 10(-5) M of 15-HPETE caused endothelial cell death in about 60% of the population: however, pretreatment of the cells with 10(-5) M of MCI-186 or concomitant addition of 10(-5) M of MCI-186 with 15-HPETE to the cultures prevented the cell death completely. These results suggest that MCI-186 may become an unique anti-ischemic drug. 相似文献
6.
Shoji Okamura Masato Kakiuchi Atsuko Sano Arasuke Nishi 《Journal of plant research》1992,105(3):503-513
Tubulin contents in the extract from cultured carrot cells at different growth phases were investigated by measuring colchicine-binding
activity. The addition of vinblastine and dithiothreitol to the reaction mixture appreciably improved the stability of both
free and colchicine-bound tubulins. Colchicine-binding activity in the cell extract obtained from stationary phase was more
labile than that from log phase though the extract showed higher affinity to colchicine. After purification, however, tubulin
from the cells at different growth phases showed the same affinity and its colchicine-binding activity was much more stable
than in crude extract. The colchicine-binding activity in the crude extract was corrected for the decay during measurement
and apparent difference in the affinity so that the activity in the cells containing different kind and amount of interefering
substances could be compared. The corrected amount of colchicine that binds to the 100,000×g extract was 46 pmol/105 cells at log phase. It decreased with the progression of culture age from linear to stationary phase. Combining the data
with the morphological observation, it was suggested that the log phase cells contained larger free tubulin pool than the
linear or stationary phase cells. 相似文献
7.
Requirement of c-kit for development of intestinal pacemaker system. 总被引:65,自引:0,他引:65
H Maeda A Yamagata S Nishikawa K Yoshinaga S Kobayashi K Nishi S Nishikawa 《Development (Cambridge, England)》1992,116(2):369-375
A discovery that the protooncogene encoding the receptor tyrosine kinase, c-kit, is allelic with the Dominant white spotting (W) locus establishes that c-kit plays a functional role in the development of three cell lineages, melanocyte, germ cell, and hematopoietic cell which are defective in W mutant mice. Recent analyses of c-kit expression in various tissues of mouse, however, have demonstrated that c-kit is expressed in more diverse tissues which are phenotypically normal in W mutant mice. Thus, whether or not c-kit expressed outside the three known cell lineages plays a functional role is one of the important questions needing answering in order to fully elucidate the role of c-kit in the development of the mouse. Here, we report that some of the cells in smooth muscle layers of developing intestine express c-kit. Blockade of its function for a few days postnatally by an antagonistic anti-c-kit monoclonal antibody (mAb) results in a severe anomaly of gut movement, which in BALB/c mice produces a lethal paralytic ileus. Physiological analysis indicates that the mechanisms required for the autonomic pacing of contraction in an isolated gut segment are defective in the anti-c-kit mAb-treated mice, W/Wv mice and even W/+ mice. These findings suggest that c-kit plays a crucial role in the development of a component of the pacemaker system that is required for the generation of autonomic gut motility. 相似文献
8.
A leptomycin B resistance gene of Schizosaccharomyces pombe encodes a protein similar to the mammalian P-glycoproteins 总被引:4,自引:0,他引:4
Kazunori Nishi Minoru Yoshida Marie Nishimura Mitsuo Nishikawa Makoto Nishiyama Sueharu Horinouchi Teruhiko Beppu 《Molecular microbiology》1992,6(6):761-769
Screening for leptomycin B (LMB)-resistant transformants in a gene library constructed in Schizosaccharomyces pombe with the chromosomal DNA of an LMB-resistant mutant of S. pombe and with multicopy plasmid pDB248' as the vector led to the isolation of a gene, named pmd1+, encoding a 1362-amino-acid protein. This protein showed great similarity in amino acid sequence to the mammalian P-glycoprotein encoded by the multidrug resistance gene, mdr, and the Saccharomyces cerevisiae a-factor transporter encoded by STE6. In addition, computer analyses predicted that the protein encoded by pmd1+ formed an intramolecular duplicated structure and each of the halves contained six transmembrane regions as well as two ATP-binding domains, as observed with the P-glycoproteins and the STE6 product. Consistent with this was that S. pombe cells containing the pmd1+ gene on a multicopy plasmid showed resistance not only to LMB but also to several cytotoxic agents. The pmd1 null mutants derived by gene disruption were viable and hypersensitive to these agents. All these data suggest that the pmd1+ gene encodes a protein that is a structural and functional counterpart of mammalian mdr proteins. 相似文献
9.
E Kita N Katsui K Nishi M Emoto Y Yanagase S Kashiba 《Journal of general microbiology》1986,132(11):3095-3103
Mice orally infected with Campylobacter jejuni developed focal infiltrative necrotic lesions in the liver, as determined by both histology and liver function tests. The initial histopathological feature was a focal infiltrative lesion in the parenchyma and portal triads. Foci of infiltrative lesions became necrotic between days 30 and 60 post-inoculation (p.i.). During this period, portal infiltrates increased in severity. From month 4 p.i., focal areas of infiltrative necrosis in the liver parenchyma became extensive. Study of liver function demonstrated mild elevations of transaminases, alkaline phosphatase and lactic dehydrogenase, and also the presence of hypoalbuminaemia. Although histopathological changes of the liver became gradually more marked after day 30 p.i., liver functions of infected mice were most affected at 2 months p.i. The capacity of C. jejuni to induce hepatic lesions seemed to be related to that of organisms to persist in the gall bladder; there was no correlation between biliary carriage in infected mice and positive faecal culture. 相似文献
10.
Macromolecules purified from Lentinus edodes mycelia cultureenhanced adventitious root formation in Azukia epicotyl cuttings.Partial purification by sequential column chromatographies gavematerial composed of 71% polysaccharides and 29% proteins. Thesugar moiety consisted of mainly xylose, glucose and arabinose,the sum of their contents being more than 76% of the total carbohydrates.The protein moiety consisted of mainly glycine and serine, whichaccounted for more than 40% of the total amino acid residues. (Received June 9, 1984; Accepted November 12, 1984) 相似文献