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21.
Protein kinase C phosphorylation of desmin at four serine residues within the non-alpha-helical head domain 总被引:12,自引:0,他引:12
S Kitamura S Ando M Shibata K Tanabe C Sato M Inagaki 《The Journal of biological chemistry》1989,264(10):5674-5678
We reported that phosphorylation by either cAMP-dependent protein kinase or protein kinase C (Ca2+/phospholipid-dependent enzyme) in vitro induces disassembly of the desmin filaments (Inagaki, M., Gonda, Y., Matsuyama, M., Nishizawa, K., Nishi, Y., and Sato, C. (1988) J. Biol. Chem. 263, 5970-5978). For this subunit protein, Ser-29, Ser-35, and Ser-50 within the non-alpha-helical head domain were shown to be the sites of phosphorylation for cAMP-dependent protein kinase (Geisler, N., and Weber, K. (1988) EMBO J. 7, 15-20). In the present work, we identified the sites of desmin phosphorylated in vitro by other protein kinase which affects the filament structure. The protein kinase C-phosphorylated desmin was hydrolyzed with trypsin, and the phosphorylated peptides were isolated by reverse-phase chromatography. Sequential analysis of the purified phosphopeptides, together with the known primary sequence, revealed that Ser-12, Ser-29, Ser-38, and Ser-56 were phosphorylated by protein kinase C. All four sites are located within the non-alpha-helical head domain of desmin. Ser-12, Ser-38, and Ser-56, specifically phosphorylated by protein kinase C, have arginine residues at the carboxyl-terminal side (Arg-14, Arg-42, and Arg-59, respectively). Ser-29 phosphorylated by both protein kinase C and cAMP-dependent protein kinase has arginine residues at the amino and carboxyl termini (Arg-27 and Arg-33). These findings support the view that the head domain-specific phosphorylation strongly influences desmin filament structure; however, each protein kinase differed with regard to site recognition on this domain. 相似文献
22.
Microsatellite DNA markers for rice chromosomes 总被引:45,自引:1,他引:44
H. Akagi Y. Yokozeki A. Inagaki T. Fujimura 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1996,93(7):1071-1077
We found 369 complete microsatellites, of which (CGG/GCC)n was the most frequent, in 11 798 rice sequences in the database. Of these microsatellites, 35 out of 45 could be successfully converted into microsatellite DNA markers using sequence information in their flanking regions. Thus, the time and labor used to develop new microsatellite DNA markers could be saved by using these published sequences. Twenty eight polymorphic markers between Asominori (japonica) and IR24 (indica) have been correctly mapped on the rice genome and microsatellites appear to be randomly distributed in the rice chromosomes. Integration of these markers with the published microsatellite DNA markers showed that about 35% of the rice chromosomes were covered by the 56 microsatellite DNA markers. These microsatellites were hypervariable and were easily to assay by PCR; they were distributed to all chromosomes and therefore, one can easily select plants carrying desired chromosome regions using these microsatellite DNA markers. Thus, microsatellite maps should aid the development of new breeds of rice saving time, labor, and money. 相似文献
23.
Hiraishi Katsuya; Suzuki Keiji; Hakomori Sen-itiroh; Adachi Masakazu 《Glycobiology》1993,3(4):381-390
Apoptosis (programmed cell death) is a basic physiological processwhich determines specific patterns of tissue size and shape,and balance of cell number, during morphogenesis, and seemsto play an integral role in oncogenic progression. Since dramaticchanges of cellular glycosylation pattern are well known tobe closely correlated with differentiation, development andoncogenesis, it is likely that similar specific changes areassociated with apoptosis. However, this possibility has notbeen systematically investigated. We therefore carried out histologicalstudies of many tumours and normal tissues for which a highincidence of apoptosis is believed to occur. Sections were stainedwith monoclonal antibodies (MoAbs) directed to carbohydrateantigens Ley and Lex, proliferating cellular nuclear antigen(PCNA) and Fas (previously claimed to be an apoptosis-inducingantigen). Antibody staining patterns were compared with morphologicalcell characteristics as revealed by haematoxylin/eosin staining,and DNA fragmentation patterns (a marker of apoptosis) as revealedby 3'-OH nick-end labelling technique. We found that expressionof Ley (defined by MoAb BM1) is closely correlated with theprocess of apoptosis, but not with cell proliferation or necrosis.Within Ley-positive areas of tissue sections, typical apoptoticmorphological changes and DNA fragmentation (as revealed bypositive nick-end labelling) were frequently observed in certainloci, although not all Ley-positive cells showed such signsof apoptosis. Ley-positive areas showed consistent negativestaining by MoAb directed to PCNA and negative or weak stainingby MoAb directed to Fas antigen, regardless of tissue source.No such trends were observed for Lex glycosylation. We concludethat Ley expression is a useful phenotypic marker predictiveof apoptosis, i.e. some (although not all) Ley-positive cellssubsequently become apoptotic. apoptosis expression glycosylation patterns Ley antigen 3'-OH nick-end labelling 相似文献
24.
S. Goto H. Nagai N. Inagaki A. Koda 《Prostaglandins, leukotrienes, and essential fatty acids》1993,48(6)
Combined treatment with propranolol and reserpine enhanced acetylcholine-induced doseresponse curves for bronchoconstriction in guinea pigs in vivo. This airway hyperreactivity model was investigated pharmacologically. (1) Increased capillary permeability and increases in leukocytes in bronchoalveolar lavage fluid (BALF) were not observed after this combined treatment. (2) The increased airway sensitivity to acetylcholine produced by propranolol and reserpine was inhibited by ketotifen and theophylline, reported in clinical studies to inhibit airway hyperreactivity. (3) Two leukotriene (LT) receptor antagonists, MCI-826 and FPL-55712, clearly inhibited this increased airway reactivity. (4) A thromboxane A2 (TXA2) receptor antagonist, ONO-3708, and TXA2 synthetase inhibitor, OKY-046, also inhibited this increased airway reactivity.These results suggest that the airway hyperreactivity model produced by propranolol and reserpine in guinea pigs is a valuable pharmacological tool for investigating a remedy and LT and TXA2 may be involved in the onset of this airway hyperreactivity. 相似文献
25.
26.
Circular DNA Plasmid in the Phytopathogenic Fungus Alternaria Alternata: Its Temperature-Dependent Curing and Association with Pathogenicity 总被引:2,自引:0,他引:2
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We found the presence of plasmid DNA in strain T88-56 of the Japanese pear pathotype of Alternaria alternata, which causes black spot of certain cultivars of Japanese pear by producing host-specific AK-toxin. The plasmid, designated pAAT56, was identified to be an ~5.4-kilobase (kb) circular molecule by electron microscopic observation and restriction endonuclease mapping. Southern blot analysis showed that pAAT56 DNA had no homology with either nuclear or mitochondrial DNA. Cultures of strain T88-56 grown at 26° showed markedly reduced plasmid levels relative to those grown at lower temperatures. The strain was completely cured of pAAT56 during growth at 29°. Temperature-dependent curing of pAAT56 was confirmed by using single-protoplast isolates from mycelia grown at 23°, most of which maintained the plasmid, and from mycelia grown at 29°, most of which had lost the plasmid. Northern blot analysis detected the presence of three RNA species (~1.7, 2.7 and 5.4 kb) transcribed from pAAT56. The biological function of pAAT56 was observed using single-protoplast isolates from mycelia that either contained or had been cured of pAAT56. The plasmid-containing isolates tended to be reduced in AK-toxin production and pathogenicity compared with the plasmid-cured isolates. 相似文献
27.
Rika Morishita Shinsuke Saga Noriko Kawamura †Yoshio Hashizume ‡Toshiaki Inagaki Kanefusa Kato Tomiko Asano 《Journal of neurochemistry》1997,68(2):820-827
Abstract: The localization of two forms of the γ subunit of G proteins, γ3 and γ12, was examined in the mammalian brain. Concentrations of these two γ subunits increased markedly, as did those of glial fibrillary acidic protein, during postnatal development in the rat cerebral cortex. In aged human brains, by contrast, the concentration of γ3 tended to decrease with age, whereas that of γ12 in the temporal cortex increased slightly. An immunohistochemical study of human brains revealed that γ3 was abundant in the neuropil, whereas γ12 was localized in glial cells. In the hippocampal formation of aged human brains, levels of γ12-positive cells, as well as levels of glial fibrillary acidic protein- and vimentin-positive astrocytes, increased, in particular in the CA1 subfield and the prosubiculum, in which there was a decrease in the number of pyramidal cells. The appearance of γ12-positive cells associated with the loss of pyramidal cells was also observed in the hippocampus of rats that had been treated with kainic acid. These results indicate that γ12 is strongly expressed in reactive astrocytes. In a study of cultured neural cells, we found that γ12 was predominant in glioma cells, such as C6 and GA-1 cells, in contrast with the specific localization of γ3 in PC12 pheochromocytoma cells, which are neuron-like cells. Taken together, the results indicate that γ3 and γ12 are selectively expressed in neuronal and glial cells, respectively, and that concentrations of γ3 and γ12 in the brain are related to the numbers and/or extent of maturation of these cells. 相似文献
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30.
Takeaki Miyamoto Hiroshi Sakabe Hiroshi Inagaki 《International journal of biological macromolecules》1983,5(3):188-190
The β-structure of S-caboxymethyl derivatives of microfibrillar proteins isolated from Merino wool was investigated by X-ray diffraction for comparison with the structur of β-keratin. The S-carboxymethylated microfibrillar proteins(SCMKA) w well-oriented β-films of SCMKA weer obtained by stretching the SCMKA cast films in steam up to about 300% extesnsion. It was found that the reflections in β-pattern of SCMKA may be indexed on a pseudo-orthorhombic unit cell with a =0.94 nm, b = 0.66 nm and c = nm, where the ab, and c axes are in the direction of the interchain hydrogen bonding, the main chain(fibre axis) and the side chain, respectively. The unit cell dimesnions evaluated for SCMKA were almost the same as those for β-keratin, suggeting that few peptide sequences containing S-carboxymethyl cystine may be involved in the formation of β-structure from SCMKA. 相似文献