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141.
Wiriyathanawudhiwong N Ohtsu I Li ZD Mori H Takagi H 《Applied microbiology and biotechnology》2009,81(5):903-913
l-Cysteine is an important amino acid in terms of its industrial applications. We previously found marked production of l-cysteine directly from glucose in recombinant Escherichia coli cells by the combination of enhancing biosynthetic activity and weakening the degradation pathway. Further improvements in
l-cysteine production are expected to use the amino acid efflux system. Here, we identified a novel gene involved in l-cysteine export using a systematic and comprehensive collection of gene-disrupted E. coli K-12 mutants (the Keio collection). Among the 3,985 nonessential gene mutants, tolC-disrupted cells showed hypersensitivity to l-cysteine relative to wild-type cells. Gene expression analysis revealed that the tolC gene encoding the outer membrane channel is essential for l-cysteine tolerance in E. coli cells. However, l-cysteine tolerance is not mediated by TolC-dependent drug efflux systems such as AcrA and AcrB. It also appears that other
outer membrane porins including OmpA and OmpF do not participate in TolC-dependent l-cysteine tolerance. When a low-copy-number plasmid carrying the tolC gene was introduced into E. coli cells with enhanced biosynthesis, weakened degradation, and improved export of l-cysteine, the transformants exhibited more l-cysteine tolerance and production than cells carrying the vector only. We concluded that TolC plays an important role in
l-cysteine tolerance probably due to its export ability and that TolC overexpression is effective for l-cysteine production in E. coli.
Natthawut Wiriyathanawudhiwong and Iwao Ohtsu contributed equally to this work. 相似文献
142.
Antonella Pepe Liang Sun Ilaria Zanardi Xinyuan Wu Cristiano Ferlini Gabriele Fontana Ezio Bombardelli Iwao Ojima 《Bioorganic & medicinal chemistry letters》2009,19(12):3300-3304
Novel C-seco-taxoids were synthesized from 10-deacetylbaccatin III and their potencies evaluated against drug-sensitive and drug-resistant cancer cell lines. The drug-resistant cell lines include ovarian cancer cell lines resistant to cisplatin, topotecan, adriamycin and paclitaxel overexpressing class III β-tubulin, A2780TC1 and A2780TC3. The last two cell lines were selected through chronic exposure of A2780wt to paclitaxel and Pgp blocker cyclosporine. All novel C-seco-taxoids exhibited remarkable potency against A2780TC1 and A2780TC3 cell lines, and no cross resistance to cisplatin- and topotecan-resistant cell lines, A2780CIS and A2780TOP. Four of those C-seco-taxoids exhibit much higher activities than IDN5390 against paclitaxel-resistant cell lines, A2780ADR, A2780TC1 and A2780TC3. SB-CST-10202 possesses the best all-round high potencies across different drug-resistant cell lines. Molecular modeling studies, including molecular dynamics simulations, on the drug-protein complexes of class I and III β-tubulins were performed to identify possible cause of the remarkable potency of these C-seco-taxoids against paclitaxel-resistant cell lines overexpressing class III β-tubulin. 相似文献
143.
144.
Naoyuki Kaneko Koh Miura Hideaki Karasawa Hiroyuki Sasaki Satoru Yokoyama Akihiro Yamamura Hiroki Nagase Iwao Sasaki 《Biochemical and biophysical research communications》2009,390(4):1235-4182
Ndc80 has been shown to play an important role in stable microtubule-kinetochore attachment, chromosome alignment, and spindle checkpoint activation in mitosis. It is composed of two heterodimers, CDCA1-KNTC2 and SPC24-SPC25. Overexpression of CDCA1 and KNTC2 is reported to be associated with poor prognosis in non-small cell lung cancers (NSCLC), and siRNA-mediated knockdown against CDCA1 or KNTC2 has been found to inhibit cell proliferation and induction of apoptosis in NSCLC, ovarian cancer, cervical cancer and glioma. Therefore, CDCA1 and KNTC2 can be considered good candidates for molecular target therapy as well as diagnosis in some cancers. However, the role of the Ndc80 complex in colorectal and gastric cancers (CRC and GC) still remains unclear. In the present study, we used qRT-PCR to evaluate the expression levels of CDCA1, KNTC2, SPC24 and SPC25 in CRC and GC and employed siRNA-mediated knockdown to examine cell proliferation and apoptosis. mRNA overexpression of these four genes was observed in CRCs and GCs when compared with the corresponding normal mucosae. Additionally, the expression levels of tumor/normal ratios of CDCA1, KNTC2, SPC24 and SPC25 correlated with each other in CRCs. MTT assays revealed that cell growths after the siRNA-mediated knockdown of either CDCA1 or KNTC2 were significantly suppressed, and flow cytometry analyses revealed significant increases of the subG1 fractions after knockdown against both genes. Our present results suggest that expressional control of component molecules of Ndc80 can be utilized for molecular target therapy of patients with CRC and GC. 相似文献
145.
Kazuhisa Nozawa Maki Fujishiro Mikiko Kawasaki Hiroshi Kaneko Kazuhisa Iwabuchi Mitsuaki Yanagida Fujihiko Suzuki Keiji Miyazawa Yoshinari Takasaki Hideoki Ogawa Kenji Takamori Iwao Sekigawa 《Arthritis research & therapy》2009,11(6):R174
Introduction
A protein analysis using a mass spectrometry indicated that there are serum proteins showing significant quantitative changes after the administration of infliximab. Among them, connective tissue growth factor (CTGF) seems to be related to the pathogenesis of rheumatoid arthritis (RA). Therefore, this study was conducted to investigate how CTGF is associated with the disease progression of RA.Methods
Serum samples were collected from RA patients in active or inactive disease stages, and before or after treatments with infliximab. CTGF production was evaluated by ELISA, RT-PCR, indirect immunofluorescence microscopy, and immunoblotting. Osteoclastogenesis was evaluated using tartrate-resistant acid phosphatase (TRAP) staining, a bone resorption assay and osteoclasts specific catalytic enzymes productions.Results
The serum concentrations of CTGF in RA were greater than in normal healthy controls and disease controls. Interestingly, those were significantly higher in active RA patients compared to inactive RA patients. Furthermore, the CTGF levels significantly were decreased by infliximab concomitant with the disease amelioration. In addition, tumour necrosis factor (TNF)α can induce the CTGF production from synovial fibroblasts even though TNFα can oppositely inhibit the production of CTGF from chondrocytes. CTGF promoted the induction of the quantitative and qualitative activities of osteoclasts in combination with M-CSF and receptor activator of NF-κB ligand (RANKL). In addition, we newly found integrin αVβ3 on the osteoclasts as a CTGF receptor.Conclusions
These results indicate that aberrant CTGF production induced by TNFα plays a central role for the abnormal osteoclastic activation in RA patients. Restoration of aberrant CTGF production may contribute to the inhibition of articular destruction in infliximab treatment. 相似文献146.
147.
148.
Toyoki Iwao Takashi Yamaguchi Akira Kurashima Miyuki Maegawa 《Phycological Research》2010,58(2):132-137
Fragments of Ecklonia cava Kjellman were cultured under controlled laboratory conditions of light irradiance, water temperature, and photoperiod. To clarify the relationship between the maturation of E. cava and the photosynthetic products, laminaran, the content in the fragments was measured with the progress of maturation. The culture conditions ranged from 12.5 to 100 µmol m?2 s?1, 10–25°C, and 14 : 10 h LD (light : dark) to 10 : 14 h LD. In the case of low light conditions, despite an optimum temperature for maturation, the fragments did not form sori and laminaran was not accumulated during the culture period. In the case of sufficient light and non‐optimum temperature conditions, the fragments did not form sori, but laminaran was accumulated. When the fragments were cultured under optimum light and temperature conditions for maturation, laminaran was accumulated in the early stage of maturation, just before or after cortex of the bladelets thickened, and decreased with the progress of maturation, and all fragments matured regardless of the length of the photoperiod. So, these results support the idea that laminaran is used as the main respiratory substrate in the maturation of E. cava. 相似文献
149.
Kanbe Y Kim MH Nishimoto M Ohtake Y Yoneya T Ohizumi I Tsunenari T Taniguchi K Kaiho S Nabuchi Y Araya H Kawata S Morikawa K Jo JC Kwon HA Lim HS Kim HY 《Bioorganic & medicinal chemistry letters》2006,16(18):4959-4964
In order to develop orally active pure antiestrogens, we incorporated the carboxy-containing side chains into the 7alpha-position of the steroid scaffold and found that 17-keto derivative CH4893237 (12b) functioned as a pure antiestrogen with its oral activity much superior to clinically used pure antiestrogen, ICI182,780. Results from the pharmacokinetic evaluation indicated that the potent antiestrogen activity at oral dosing in mice attributed to both improved absorption from the intestinal wall and metabolic stability in liver. 相似文献
150.
Zhou J Huang B Suzuki H Fujisaki K Igarashi I Xuan X 《The Journal of parasitology》2006,92(1):208-210
Actin is a ubiquitous and highly conserved microfilament protein that is hypothesized to play a mechanical force-generating role in the unusual gliding motility of sporozoan zoites and their active penetration of host cells. We have identified and isolated an actin gene from a Babesia gibsoni cDNA library by random sequencing. The complete nucleotide sequence of the actin gene is 1,243 bp; a single open reading frame encodes a polypeptide of 377 amino acid residues. The deduced amino acid sequence showed a high homology with actins from other species, especially with reported apicomplexan protozoans. The antiserum against recombinant actin expressed in Escherichia coli recognizes a 42-kDa native protein, which is consistent with its expected size. Immunofluorescence and confocal microscopic observation revealed that the protein is diffusely distributed throughout the B. gibsoni parasites. 相似文献