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91.
A symposium celebrating the 40th anniversary of the Protein Data Bank archive (PDB), organized by the Worldwide Protein Data Bank, was held at Cold Spring Harbor Laboratory (CSHL) October 28-30, 2011. PDB40's distinguished speakers highlighted four decades of innovation in structural biology, from the early?era of structural determination to future directions for the field. 相似文献
92.
Moue M Tohno M Shimazu T Kido T Aso H Saito T Kitazawa H 《Biochimica et biophysica acta》2008,1780(2):134-144
To study the immune responses of porcine intestinal epithelial cells to gram-negative bacteria via toll-like receptors (TLRs), originally established porcine intestinal epitheliocyte (PIE) cells were treated with lipopolysaccharide (LPS) or swine-specific enterotoxigenic Escherichia coli (ETEC). Real-time quantitative PCR revealed that PIE cells expressed TLR1-9 and MD-2 mRNAs, preferentially expressed TLR4/MD-2. Immunostaining of PIE cells revealed that TLR4 was precisely expressed in PIE cells at the protein level. PIE cells treated with LPS had up-regulated expression of several TLRs (TLR2, 3, 4, 5 and 8), type 1 helper T (Th1) cytokines (interleukin (IL)-1alpha, IL-1beta, IL-6, IL-15, 18, leukemia inhibitory factor (LIF), and interferon (IFN)-beta), and chemokines (monocyte chemoattractant protein (MCP)-1 and IL-8). ETEC enhanced the expression of TLR2, Th1 type cytokines (IL-1alpha, IL-12p35 and IL-6) and chemokines (MCP-1 and IL-8). These results indicate that PIE induces inflammatory responses by up-regulating Th1 cytokines and chemokines in response to LPS or ETEC, suggesting that PIE is a useful cell line for studying inflammatory responses via TLR4/MD-2 in intestinal epithelial cells. 相似文献
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We investigated the ability of Enteromorpha intestinalis (L.) Link to take up pulses of different species of nitrogen simultaneously, as this would be an important mechanism to enhance bloom ability in estuaries. Uptake rates and preference for NH4+ or NO3− following 1, 3, 6, 9, 12 or 24 h of exposure to either 15NH4NO3 or NH415NO3 were determined by disappearance of N from the medium. Differences in assimilation rates for NH4+ or NO3− were quantified by the accumulation of NH4+, NO3−, and atom % 15N in the algal tissue. NH4+ concentration was reduced more quickly than water NO3− concentration. Water column NH4+ concentration after the longest time interval was reduced from 300 to 50 μM. Water NO3− was reduced from 300 to 150 μM. The presence of 15N or 14N had no effect on uptake of either NH4+ or NO3−. 15N was removed from the water at an almost identical rate and magnitude as 14N. Differences in accumulation of 15NH4+ and 15NO3− in the tissue reflected disappearance from the water; 15N from NH4+ accumulated faster and reached an atom % twice that of 15N from NO3−. This outcome suggested that when NH4+ and NO3− were supplied in equal concentrations, more NH4+ was taken up and assimilated. The ability to take up high concentrations of NH4+, and NO3− simultaneously is important for bloom-forming species of estuarine macroalgae subject to multiple nutrient species from various sources. 相似文献
96.
Yutaka Suzuki Mitsuru Haruki Kazufumi Takano Masaaki Morikawa Shigenori Kanaya 《European journal of biochemistry》2004,271(7):1372-1381
A psychrotrophic bacterium Shewanella sp. strain SIB1 was grown at 4 and 20 degrees C, and total soluble proteins extracted from the cells were analyzed by two-dimensional polyacrylamide gel electrophoresis. Comparison of these patterns showed that the cellular content of a protein with a molecular mass of 28 kDa and an isoelectric point of four greatly increased at 4 degrees C compared to that at 20 degrees C. Determination of the N-terminal amino acid sequence, followed by the cloning and sequencing of the gene encoding this protein, revealed that this protein is a member of the FKBP family of proteins with an amino acid sequence identity of 56% to Escherichia coli FKBP22. This protein was overproduced in E. coli in a His-tagged form, purified, and analyzed for peptidyl-prolyl cis-trans isomerase activity. When this activity was determined by the protease coupling assay using N-succinyl-Ala-Leu-Pro-Phe-p-nitroanilide as a substrate at various temperatures, the protein exhibited the highest activity at 10 degrees C with a k(cat)/K(m) value of 0.87 micro m(-1) x s(-1). When the peptidyl-prolyl cis-trans isomerase activity was determined by the RNase T(1) refolding assay at 10 and 20 degrees C, the protein exhibited higher activity at 10 degrees C with a k(cat)/K(m) value of 0.50 micro m(-1) x s(-1). These k(cat)/K(m) values are lower but comparable to those of E. coli FKBP22. We propose that a FKBP family protein is involved in cold-adaptation of psychrotrophic bacteria. 相似文献
97.
Tomoki Matsuda Makoto Fujikawa Mitsuru Haruki Xiao-Feng Tang Satoshi Ezaki Tadayuki Imanaka Masaaki Morikawa Shigenori Kanaya 《Extremophiles : life under extreme conditions》2001,5(3):177-182
Interactions of TBP-interacting protein (TIP26), TBP, and TFB from a hyperthermophilic archaeon Thermococcus kodakaraensis KOD1 with TATA-DNA were examined by electrophoretic mobility shift assay. Tk-TFB formed a ternary complex with Tk-TBP and TATA-DNA. Tk-TIP26 did not inhibit the formation of this ternary complex, but interacted with it to form a TIP26/TFB/TBP/DNA quaternary complex. This interaction is rather weak, and a large excess of Tk-TIP26 over Tk-TBP is required to fully convert the TFB/TBP/DNA ternary complex to the quaternary complex. However, determination of the concentration of Tk-TIP26 and Tk-TBP in KOD1 cells by Western blotting analysis indicated that the concentration of Tk-TIP26 is approximately ten times that of Tk-TBP, suggesting that the quaternary complex might also form in vivo. 相似文献
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99.
Active Subtilisin-Like Protease from a Hyperthermophilic Archaeon in a Form with a Putative Prosequence 总被引:6,自引:2,他引:4
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Yuji Kannan Yuichi Koga Yohei Inoue Mitsuru Haruki Masahiro Takagi Tadayuki Imanaka Masaaki Morikawa Shigenori Kanaya 《Applied microbiology》2001,67(6):2445-2452
The gene encoding subtilisin-like protease T. kodakaraensis subtilisin was cloned from a hyperthermophilic archaeon Thermococcus kodakaraensis KOD1. T. kodakaraensis subtilisin is a member of the subtilisin family and composed of 422 amino acid residues with a molecular weight of 43,783. It consists of a putative presequence, prosequence, and catalytic domain. Like bacterial subtilisins, T. kodakaraensis subtilisin was overproduced in Escherichia coli in a form with a putative prosequence in inclusion bodies, solubilized in the presence of 8 M urea, and refolded and converted to an active molecule. However, unlike bacterial subtilisins, in which the prosequence was removed from the catalytic domain by autoprocessing upon refolding, T. kodakaraensis subtilisin was refolded in a form with a putative prosequence. This refolded protein of recombinant T. kodakaraensis subtilisin which is composed of 398 amino acid residues (Gly−82 to Gly316), was purified to give a single band on a sodium dodecyl sulfate (SDS)-polyacrylamide gel and characterized for biochemical and enzymatic properties. The good agreement of the molecular weights estimated by SDS-polyacrylamide gel electrophoresis (44,000) and gel filtration (40,000) suggests that T. kodakaraensis subtilisin exists in a monomeric form. T. kodakaraensis subtilisin hydrolyzed the synthetic substrate N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide only in the presence of the Ca2+ ion with an optimal pH and temperature of pH 9.5 and 80°C. Like bacterial subtilisins, it showed a broad substrate specificity, with a preference for aromatic or large nonpolar P1 substrate residues. However, it was much more stable than bacterial subtilisins against heat inactivation and lost activity with half-lives of >60 min at 80°C, 20 min at 90°C, and 7 min at 100°C. 相似文献
100.
Haruki Takahashi 《Mycoscience》2001,42(4):347-353
Three new species of Agaricales are described and illustrated from eastern Honshu, Japan:Boletus rhodocarpus sp. nov. (sectionLuridi), forming large, deep red basidiomata with a pileus covered with small, blackish brown scales, was found on ground in a highland
forest dominated byTsuga diversifolia andAbies veitchii; Phaeomarasmius laccarioides sp. nov. (subgenusCarpophilus), forming a squamulose-fibrillose, reddish brown pileus in which the pileipellis consists of chains of thick-walled sphaerocysts
with heavily incrusting, brown pigment, was found on a fallen fruit ofLiquidambar styraciflua; Pluteus phaeocephalus sp. nov. (subsectionHispidodermini of sectionCelluloderma), forming a dark brown, velvety pileus and a white stipe densely covered with dark brown punctate scales, was found on dead
fallen twigs ofQuercus serrata. 相似文献