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31.
32.
High molecular weight binding components which bind [125I] mouse β nerve growth factor exist in human serum. The binding of β nerve growth factor to the serum components was inhibited at alkaline condition. After gel filtration of human serum on a Sephadex G-150 column at neutral condition, the nerve growth factor-like immunoreactivity was observed in only one peak, differing from the high molecular weight serum components. However, at alkaline condition two peaks with nerve growth factor-like immunoreactivity appeared; one was almost at the position observed at neutral pH, and the other was a new peak eluted approximately to the column volume. these results suggest that there are at least two nerve growth factor-like molecules in human serum and most of the nerve growth factor in the serum exists in a complex form associated with serum components with high molecular weight. 相似文献
33.
Toshiya Endo Masanao Oya Kyozo Hayashi Tatsuo Miyazawa 《Journal of Protein Chemistry》1989,8(4):575-581
Temperature dependence of proton nuclear magnetic resonance spectra has been followed for cobrotoxin, a postsynaptic neurotoxin fromNaja naja atra venom. Several aromatic amino-acid residues, including the functionally essential Trp-29 located at the tip of the central loop of the molecule, have been found to undergo a thermal structural transition above the global thermal denaturation temperature. It is suggested that a local structure around these residues behaves somehow independently of the rest of the molecule, and that such structural organization may be favorable for a conformational change of a neurotoxin molecule on binding to acetylcholine receptor. 相似文献
34.
Single blastomeres from eight-cell stage bovine embryos matured and fertilized in vitro were electrically fused with enucleated oocytes matured in vitro. In experiment 1, The percentage of these reconstituted embryos developed to the two- to eight-cell stage 48 hr after electrofusion was increased when both the eight-cell embryos and the enucleated oocytes were derived from oocytes cultured with granulosa cells (14% vs. 38%). In experiment 2, the relationship between activation of oocytes and developmental ability of reconstituted embryos was examined. Although both ethanol and electrical stimulation efficiently induced parthenogenetic activation of oocytes matured in vitro for 26–28 hr (ethanol, 89%; electrical stimulation, 73%), the ratio of the second polarbody extrusion differed (80% vs. 22%). Ethanol-treated enucleated oocytes, however, were not significantly different from the early cleavage of the reconstituted embryos 48 hr after electrofusion (nontreated, 38%; treated, 43%). In experiment 3, reconstituted embryos at the two- to eight-cell stage 48 hr after the electrofusion were cocultured with granulosa cells for 6–7 days. Of 69 embryos, one developed to a morula and three developed to blastocysts. © 1993 Wiley-Liss, Inc. 相似文献
35.
Masanori Nishida Masataka Okamoto Ai Ohno Kohji Okumura Kyozo Hayashi Kiyoshi Ikeda Seiji Inoue 《Biochimica et Biophysica Acta - Proteins and Proteomics》2010,1804(11):2121-2127
α-type phospholipase A2 inhibitory protein (PLIα) isolated from the serum of the venomous snake Glyoidius brevicaudus, GbPLIα, is a homotrimer of subunits having a C-type lectin-like domain. The serum protein from nonvenomous snake Elaphe quadrivirgata, EqPLIα-LP, is homologous to GbPLIα, but it does not show any inhibitory activity against PLA2s. When a mixture of denaturant-treated monomeric forms of GbPLIα and EqPLIα-LP was used to reconstitute their trimers, no significant amounts of heterotrimers composed of GbPLIα and EqPLIα-LP subunits could be formed. On the other hand, when a mixture of denaturant-treated monomeric forms of GbPLIα and the recombinant chimeric EqPLIα-LP, Eq13Gb37Eq, in which the residues 13–36 were replaced by those of GbPLIα, was used to reconstitute their trimers, significant amounts of their heterotrimers were observed. Furthermore, when a mixture of denaturant-treated monomeric forms of EqPLIα-LP and the recombinant chimeric GbPLIα, Gb13Eq37Gb, in which the residues 13–36 were replaced by those of EqPLIα-LP, was used, significant amounts of their heterotrimers were observed. By comparison of the respective inhibitory activities of the heterotrimeric subspecies, it was suggested that the inhibitory activity of the trimer was governed by one subunit with the highest activity, and not affected by the number of these subunits. The intermolecular electrostatic interactions between Glu23 and Lys28 of GbPLIα were also suggested to be important in stabilizing the trimeric structure. The importance of the electrostatic interaction was supported by the less stability of the homotrimeric structure of a mutant GbPLIα with a single amino acid substitution, GbPLIα(K28E). 相似文献
36.
Tsuyoshi Horiguchi Fumie Sasaki Hideki Takahama Kyozo Watanabe 《Biotechnic & histochemistry》1984,59(3):143-148
Anuran tadpole tail muscle was stained en bloc by a modified light microscope silver stain for light microscopy and freeze-fractured in liquid nitrogen after partial dehydration with ethanol. The fractured specimens were observed in both secondary electron and backscattered electron modes in a scanning electron microscope. Since the cell nuclei specifically stained with silver provided high contrast against the unstained background due to atomic number contrast of backscattered electron image, various cells were easily identified by a comparison of secondary electron images and compositional images of backscattered electron signals. 相似文献
37.
A hybridoma secreting human monoclonal antibody (MAB) against Pseudomonas aeruginosa exotoxin A (PEA) was constructed by fusing Epstein-Barr virus-transformed peripheral blood lymphocytes with human B lymphoblastoid cell line TAW-925. The human-human hybridoma stably produced human IgG2 MAB at the rate of 0.4–0.5 g/ml per 106 cells per day for more than six months, and the MAB was capable of neutralizing the in vitro cytotoxic and in vivo lethal effects of PEA with approximately 100-and 70-fold, respectively, higher activity than serum polyclonal antibody preparations.Abbreviations MAB
Monoclonal Antibody
- PEA
Pseudomonas aeruginosa exotoxin A
- LPS
Lipopolysaccharides
- OMP
Outer Membrane Proteins
-
P.
Pseudomonas
- EBV
Epstein-Barr Virus
- PEG
Polyethylene Glycol 相似文献
38.
Rajaraman G Wang GQ Yan J Jiang P Gong Y Burczynski FJ 《Molecular and cellular biochemistry》2007,295(1-2):27-34
The presence of cysteine and methionine groups together with an ability to bind long-chain fatty acid (LCFA) oxidation products
makes liver fatty acid binding protein (L-FABP) an attractive candidate against hepatocellular oxidative stress. In this report,
we show that pharmacological treatment directed at modulating L-FABP level affected hepatocellular oxidant status. L-FABP
expressing 1548-hepatoma cells, treated with dexamethasone or clofibrate, decreased and increased intracellular L-FABP levels,
respectively. Oxidative stress was induced by H2O2 incubation or hypoxia–reoxygenation. The fluorescent marker, dichlorofluorescein (DCF), was employed to measure intracellular
reactive oxygen species (ROS). Hepatocellular damage was assessed by lactate dehydrogenase (LDH) level. Dexamethasone treatment
resulted in a significant increase in DCF fluorescence with higher LDH release compared to control cells. Clofibrate treatment,
however, resulted in a significant decrease in both parameters (p < 0.05). Drug treatments did not affect cytosolic activites of glutathione peroxidase (GPx), superoxide dismutase (SOD), or
catalase suggesting that the differences between treated and control cells may likely be associated with varying L-FABP levels.
We conclude that L-FABP may act as an effective endogenous cytoprotectant against hepatocellular oxidative stress. 相似文献
39.
Tea catechins have strong bitterness and influence the taste of tea. Among the 25 human bitter-taste receptors (TAS2Rs), we found that hTAS2R14 responded to catechins, in addition to hTAS2R39, a known catechin receptor. Although hTAS2R14 responded to (?)-epicatechin gallate and (?)-epigallocatechin gallate, it did not respond to (?)-epicatechin and (?)-epigallocatechin. 相似文献
40.
Shoei Furukawa Hiroshi Nishitani Kyozo Hayashi 《Biochemical and biophysical research communications》1977,76(4):1202-1206
To examine the relationship between NGF activity and neurologic disorders, the level of NGF activity was studied in the submaxillary glands of both normal and dystrophic mice. The level of NGF activity in the mice was remarkably lower than that observed in normal mice. The data suggest that NGF activity may have some relation to the cause of the disease. 相似文献