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11.
Protein translocation into Escherichia coli membrane vesicles is inhibited by functional synthetic signal peptides 总被引:7,自引:0,他引:7
A synthetic peptide corresponding to the signal sequence of wild type Escherichia coli lambda-receptor protein (LamB) inhibits in vitro translocation of precursors of both alkaline phosphatase and outer membrane protein A into E. coli membrane vesicles (half-maximal inhibition at 1-2 microM). By contrast, the inhibitory effect was nearly absent in a synthetic peptide corresponding to the signal sequence from a mutant strain that harbors a deletion mutation in the LamB signal region and displays an export-defective phenotype for this protein in vivo. Two peptides derived from pseudorevertant strains that arose from the deletion mutant and exported LamB in vivo were found to inhibit in vitro translocation with effectiveness that correlated with their in vivo export ability. Controls indicated that these synthetic signal peptides did not disrupt the E. coli membrane vesicles. These results can be interpreted to indicate that the presequences of exported proteins interact specifically with a receptor either in the E. coli inner membrane or in the cytoplasmic fraction. However, biophysical data for the family of signal peptides studied here reveal that they will spontaneously insert into a lipid membrane at concentrations comparable to those that cause inhibition. Hence, an indirect effect mediated by the lipid bilayer of the membrane must be considered. 相似文献
12.
Synergism between eosinophil cationic protein and oxygen metabolites in killing of schistosomula of Schistosoma mansoni 总被引:4,自引:0,他引:4
M Yazdanbakhsh P C Tai C J Spry G J Gleich D Roos 《Journal of immunology (Baltimore, Md. : 1950)》1987,138(10):3443-3447
To study the cytotoxic reactions responsible for mediating eosinophil damage to schistosomula of Schistosoma mansoni, we have used cytoplasts (eosinophil or neutrophil vesicles devoid of granules and nuclei, with an intact oxidase in their plasma membrane) in combination with purified eosinophil cationic protein (ECP) or major basic protein (MBP) in a cytotoxicity test toward schistosomula. Suboptimal concentrations of ECP (10(-6) M) or MBP (10(-6) M) resulting in less than 10% killing were used in combination with cytoplasts. Cytoplasts alone in the presence of immune serum tested over a wide range of cytoplast:schistosomula ratios generated superoxide and hydrogen peroxide, but were unable to damage schistosomula. However, when a suboptimal ECP concentration (10(-6) M) was combined with neutroplasts or eosinoplasts, 43.9% +/- 8.5 (n = 7) and 24.7% +/- 9.8 (n = 3), respectively, of the schistosomula were killed. Oxygen metabolites were responsible for the synergism, because cytoplasts from a patient with chronic granulomatous disease were unable to act in synergy with ECP. In contrast to ECP, no synergism was found between cytoplasts and MBP (10(-6) to 2 X 10(-5)M). These results show that oxygen metabolites are important for the killing of schistosomula by lowering the concentration of ECP needed to inflict damage. 相似文献
13.
Stimulations of arachidonate release and inositol-1,4,5-triphosphate formation are mediated by distinct G-proteins in human platelets 总被引:4,自引:0,他引:4
Addition of fluoroaluminate to human platelet suspension stimulated thromboxane synthesis and inositol-1,4,5-triphosphate formation in a time and dose dependent manner. Neomycin inhibited markedly fluoroaluminate induced inositol-1,4,5-triphosphate formation without significantly affecting thromboxane synthesis. Preincubation of platelets with PGE1, also inhibited significantly inositol-1,4,5-triphosphate formation with modest reduction of thromboxane synthesis. On the contrary, pretreatment of platelets with pertussis toxin inhibited fluoroaluminate stimulated thromboxane synthesis without affecting inositol-1,4,5-triphosphate formation. Similarly, preincubation of platelets with phorbol ester, PMA, inhibited markedly thromboxane synthesis with modest reduction of inositol-1,4,5-triphosphate formation. These results indicate that inositol-1,4,5-triphosphate formation and arachidonate release and thromboxane synthesis are controlled separately and are mediated by different G-proteins which are coupled to phospholipase C and phospholipase A2 respectively in platelets. 相似文献
14.
A radioimmunoassay for human pro-luteinizing hormone-releasing factor [pro-LRF(14-69)OH] 总被引:1,自引:0,他引:1
A Motoyama I Wakabayashi S Minami H Sugihara F Takahashi S Akira N Ling 《Endocrinologia japonica》1987,34(1):133-137
A radioimmunoassay (RIA) for human pro-LRF(14-69)OH was developed with an antiserum, generated in a rabbit, to [Tyr67]pro-LRF(47-67)NH2 conjugated to BSA. This antiserum bound 28-32% of [125I]pro-LRF(14-69)OH at a final dilution of 1:2500 and the binding was inhibited by pro-LRF(14-69)OH in a dose-dependent manner. The sensitivity of the RIA was 31.2-62.5 pg and the dose that inhibited 50% of the binding to the tracer was 280-320 pg. Intra- and inter-assay coefficients of variation at 50% inhibition were 8 and 12%, respectively. Neither LRF nor pro-LRF(14-37)OH was recognized by the antiserum. The dilution curve generated with human hypothalamic extract was parallel to that of pro-LRF(14-69)OH. In addition the extract yielded a major immunoreactive peak emerging in elution volumes concordant with [125I]pro-LRF(14-69)OH on Sephadex G-50 chromatography. 相似文献
15.
Komatsuna (Brassica campestris L. var. rapa) plants were grownhydroponically under various conditions with respect to thesupply of nitrate, and the variations in levels of natural 15Nabundance (15N) in nitrogenous fractions of leaf blades, petiolesplus midribs, and roots in these plants were analyzed. The fractionation of nitrogen isotopes during uptake of nitratewas null irrespective of the concentrations of nitrate in theculture medium. The roots had lower 15N values than that inthe nitrate applied to plants. The nitrate in the three tissuesexamined had higher 15N values than that in the nitrate applied:the values were highest in the leaf blades which were presumedto have highest activities in terms of reduction of nitrate.In contrast, the amino acids and residual fractions had lower15N values than those in the nitrate applied. These resultssuggest that reduction of nitrate is a critical step in thefractionation of nitrogen isotopes in plant tissues in vivo.
1Permanent address: Fukuoka Agricultural Experiment Station,Yoshiki 587, Chikushino, 818 Japan. (Received March 10, 1989; Accepted July 10, 1989) 相似文献
16.
Quantitative Analysis of Endogenous Gibberellins in Normal and Dwarf Cultivars of Rice 总被引:7,自引:0,他引:7
Kobayashi Masatomo; Sakurai Akira; Saka Hitoshi; Takahashi Nobutaka 《Plant & cell physiology》1989,30(7):963-969
Endogenous levels of gibberellins in shoots and ears of twodwarf rice (Oryza sativa L.) cultivars, Tan-ginbozu (dx mutant)and Waito-C (dy mutant), were analyzed and compared with thoseof normal rice cultivar, Nihonbare. The endogenous levels of13-hydroxylated gibberellins in Tan-ginbozu were much lowerthan those in Nihonbare. In Waito-C, the levels of GA19 andGA20 in the shoots were higher but that of GA1 was lower thanthe levels of these gibberellins in Nihonbare. These resultssupport the hypothesis that the dy gene controls the 3ß-hydroxylationof GA20 to GA1 while the dx gene controls a much earlier stepin the gibberellin biosynthesis. Our results indicate that GA1is the active gibberellin that regulates the vegetative growthof rice. The endogenous levels of GA4 in the ears of the twodwarf cultivars of rice were higher than the level of GA4 inthe ears of the normal cultivar, Nihonbare suggesting that thebiosynthesis of gibberellin is not blocked in the anthers ofthe dwarf rice. (Received April 27, 1989; Accepted July 12, 1989) 相似文献
17.
Conformational change of bovine serum albumin by heat treatment 总被引:1,自引:0,他引:1
Kunio Takeda Akira Wada Kazuo Yamamoto Yoshiko Moriyama Koichiro Aoki 《Journal of Protein Chemistry》1989,8(5):653-659
The thermal denaturation of bovine serum albumin (BSA) was studied at pH 2.8 and 7.0 in the range of 2–65°C. The relative proportions of -helix, -structure, and disordered structure in the protein conformation were determined as a function of temperature, by the curve-fitting method of circular dichroism spectra. With the rise of temperature at pH 7.0, the proportion of -helix decreased above 30°C and those of -structure and disordered structure increased in the same temperature range. The structural change was reversible in the temperature range below 45°C. However, the structural change was partially reversible upon cooling to room temperature subsequent to heating at 65°C. On the other hand, the structural change of BSA at pH 2.3 was completely reversible in the temperature range of 2–65°C, probably because the interactions between domains and between subdomains might disappear due to the acid expansion. The secondary structure of disulfide bridges-cleaved BSA remained unchanged during the heat treatment up to 65°C at pH 2.8 and 7.0. 相似文献
18.
Kenzo Yamanaka Akira Hasegawa Ryoji Sawamura Shoji Okada 《Biological trace element research》1989,21(1):413-417
DNA damage induced by administration of dimethylarsinic acid (DMAA) to rats and mice was investigated. At 12 h after administration of DMAA, DNA single-strand breaks were induced markedly in lung. The majority of dimethylarsine, one of the main metabolites, in the expired air was excreted within 6–18 h after administration of DMAA to rats. In vitro experiments using nuclei isolated from lung of mice indicated that DNA strand breaks were caused by dimethylarsine. Furthermore, the strand breaks after exposure to dimethylarsine were reduced in the presence of catalase and/or superoxide dismutase. These results strongly suggest that the strand breaks are induced not by dimethylarsine itself but by active oxygen, e.g., O 2 ? and ·OH, produced both by dimethylarsine and molecular oxygen. When DNA was exposed to dimethylarsine, thiobarbituric acid (TBA)-reactive intermediates andcis-thymine glycol were produced. Dimethylarsine appears to induce DNA damage by the mechanism similar to the damage produced by ionizing radiation. 相似文献
19.
Hiroshi Yonemitsu Koichiro Shimomura Motoyoshi Satake Shunji Mochida Masahiko Tanaka Thoru Endo Akira Kaji 《Plant cell reports》1990,9(6):307-310
Hairy roots were obtained following inoculation of the stems of Lobelia inflata L. with Agrobacterium rhizogenes strain ATCC 15834. These hairy roots contained agropine and mannopine. In addition, lobeline was detected by HPLC and confirmed by mass spectrometry. Various media were tested for the growth of hairy roots as well as for the content of lobeline in hairy roots. The growth rate of hairy roots cultured in Nitsch and Nitsch's medium was approximately one third of those cultured in other media. The lobeline content of hairy roots (18–54 g/g dry weight) cultured in these media was the same order of magnitude compared with that of roots of L. inflata (24 g/g dry weight) cultivated in pots. The hairy roots cultured in Nitsch and Nitsch's medium were morphologically different from those cultured in other media.Abbreviations MS medium
Murashige and Skoog's medium
- 1/2 MS medium
one-half strength of the standard Murashige and Skoog's medium
- B5 medium
Gamborg's B5 medium
- NN medium
Nitsch and Nitsch's medium
- FW
fresh weight
- DW
dry weight 相似文献
20.
Nobutaka Fujii Akira Otaka Susumu Funakoshi Toshihiro Watanabe Hiromitsu Arai Kiyoshi Bessho Haruaki Yajima 《Journal of Protein Chemistry》1988,7(2):151-156
Treatment of a mixture of Cys(R)(O) and Cys(R) with an acid was found to generate cystine in fairly good yields, when suitable R, R, and an acid were selected. An unsymmetrical cystine peptide was prepared by treatment of a mixture of Z(OMe)-Cys(R) (0)-Ala-NH2 (R=Acm or MBzl) and Z(OMe)-Cys(MBzl)-Gly-OBzl with TFA or 1 M TFMSA/TFA.3 Oxytocin was obtained in an excellent yield by TFA treatment of the protected peptide containing Cys(Acm)(0) and Cys(MBzl). Thus, formation of the disulfide bond was found feasible at the position of Cys(R) (0).The following abbreviations are used Boc
t-butyloxycarbonyl
- Z(OMe)
p-methoxybenzyloxycarbonyl
- MBzl
p-methoxybenzyl
- Acm
acetamidomethyl
- Bzl
benzyl
- Ad
l-adamantyl
- tBu
t-butyl
- TFA
trifluoroacetic acid
- TFMSA
trifluoromethanesulfonic acid
- TMSOTf
trimethylsilyl trifluoromethane sulfonate 相似文献