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91.
Fukamachi H Nakano Y Okano S Shibata Y Abiko Y Yamashita Y 《Biochemical and biophysical research communications》2005,331(1):127-131
Methyl mercaptan is derived from l-methionine by the action of l-methionine-alpha-deamino-gamma-mercaptomethane lyase (METase) and is a major component of oral malodor. This compound is highly toxic and is thought to play an important role in periodontal disease. We found that Treponema denticola, a member of the subgingival biofilm at periodontal disease sites, produced a large amount of methyl mercaptan even at low concentration of l-methionine. METase activity in a cell-free extract from T. denticola was detected by two-dimensional electrophoresis under non-denaturing conditions, and the protein spot that exhibited high METase activity was identified using a matrix-assisted laser desorption/ionization time-of-flight mass spectrometer. The identified gene produced a METase with a K(m) value for l-methionine (0.55mM) that is much lower than those of METases previously identified in the other organisms. This result suggests that T. denticola is an important producer of methyl mercaptan in the subgingival biofilm. 相似文献
92.
Kamao M Tatematsu S Hatakeyama S Sakaki T Sawada N Inouye K Ozono K Kubodera N Reddy GS Okano T 《The Journal of biological chemistry》2004,279(16):15897-15907
93.
Sawada N Kusudo T Sakaki T Hatakeyama S Hanada M Abe D Kamao M Okano T Ohta M Inouye K 《Biochemistry》2004,43(15):4530-4537
Our previous study revealed that human CYP24A1 catalyzes a remarkable metabolism consisting of both C-23 and C-24 hydroxylation pathways that used both 25(OH)D(3) and 1alpha,25(OH)(2)D(3) as substrates, while rat CYP24A1 showed extreme predominance of the C-24 over C-23 hydroxylation pathway [Sakaki, T., Sawada, N., Komai, K., Shiozawa, S., Yamada, S., Yamamoto, K., Ohyama, Y. and Inouye, K. (2000) Eur. J. Biochem. 267, 6158-6165]. In this study, by using the Escherichia coli expression system for human CYP24A1, we identified 25,26,27-trinor-23-ene-D(3) and 25,26,27-trinor-23-ene-1alpha(OH)D(3) as novel metabolites of 25(OH)D(3) and 1alpha,25(OH)(2)D(3), respectively. These metabolites appear to be closely related to the C-23 hydroxylation pathway, because human CYP24A1 produces much more of these metabolites than does rat CYP24A1. We propose that the C(24)-C(25) bond cleavage occurs by a unique reaction mechanism including radical rearrangement. Namely, after hydrogen abstraction of the C-23 position of 1alpha,25(OH)(2)D(3), part of the substrate-radical intermediate is converted into 25,26,27-trinor-23-ene-1alpha(OH)D(3), while a major part of them is converted into 1alpha,23,25(OH)(3)D(3). Because the C(24)-C(25) bond cleavage abolishes the binding affinity of 1alpha,25(OH)D(3) for the vitamin D receptor, this reaction is quite effective for inactivation of 1alpha,25(OH)D(3). 相似文献
94.
The RING finger protein, RNF8, interacts with retinoid X receptor alpha and enhances its transcription-stimulating activity 总被引:3,自引:0,他引:3
95.
Inaba Y Tian QB Okano A Zhang JP Sakagami H Miyazawa S Li W Komiyama A Inokuchi K Kondo H Suzuki T 《Journal of neurochemistry》2004,89(6):1347-1357
We cloned from a rat brain cDNA library a novel cDNA and named it a potential synaptic guanine nucleotide exchange factor (GEF) for Arf (synArfGEF (Po)) (GenBank Accession no. AB057643) based on its domain structure and localization. The cloned gene was 7410 bases long with a 3585-bp coding sequence encoding a protein of 1194 amino acids. The deduced protein contained a coiled-coil structure in the N-terminal portion followed by Sec7 and Plekstrin homology (PH) domains. Thus, the protein was a member of the Sec7 family of proteins, GEFs. Conservation of the ADP-ribosylation factor (Arf)-binding sequence suggested that the protein was a GEF for Arf. The gene was expressed specifically in the brain, where it exhibited region-specific expression. The protein was highly enriched in the postsynaptic density (PSD) fraction prepared from the rat forebrain. Uniquely, the protein interacted with PSD-95, SAP97 and Homer/Vesl 1/PSD-Zip45 via its C-terminal PDZ-binding motif and co-localized with these proteins in cultured cortical neurons. These results supported its localization in the PSD. The postsynaptic localization was also supported by immunohistochemical examination of the rat brain. The mRNA for the synArfGEF was also localized to dendrites, as well as somas, of neuronal cells. Thus, both the mRNA and the protein were localized in the postsynaptic compartments. These results suggest a postsynaptic role of synArfGEF in the brain. 相似文献
96.
Shibata S Okano S Yonemitsu Y Onimaru M Sata S Nagata-Takeshita H Inoue M Zhu T Hasegawa M Moroi Y Furue M Sueishi K 《Journal of immunology (Baltimore, Md. : 1950)》2006,177(6):3564-3576
Dendritic cell (DC)-based cancer immunotherapy has been paid much attention as a new and cancer cell-specific therapeutic in the last decade; however, little clinical outcome has been reported. Current limitations of DC-based cancer immunotherapy include sparse information about which DC phenotype should be administered. We here report a unique, representative, and powerful method to activate DCs, namely recombinant Sendai virus-modified DCs (SeV/DC), for cancer immunotherapy. In vitro treatment of SeV without any bioactive gene solely led DCs to a mature phenotype. Even though the expression of surface markers for DC activation ex vivo did not always reach the level attained by an optimized amount of LPS, superior antitumor effects to B16F1 melanoma, namely tumor elimination and survival, were obtained with use of SeV-GFP/DC as compared with those seen with LPS/DC in vivo, and the effect was enhanced by SeV/DC-expressing IFN-beta (SeV-murine IFN-beta (mIFN-beta)/DC). In case of the treatment of an established tumor of B16F10 (7-9 mm in diameter), a highly malignant subline of B16 melanoma, SeV-modified DCs (both SeV-GFP/DC and SeV-mIFN-beta/DC), but not immature DC and LPS/DC, dramatically improved the survival of animals. Furthermore, SeV-mIFN-beta/DC but not other DCs could lead B16F10 tumor to the dormancy, associated with strongly enhanced CD8+ CTL responses. These results indicate that rSeV is a new and powerful tool as an immune booster for DC-based cancer immunotherapy that can be significantly modified by IFN-beta, and SeV/DC, therefore, warrants further investigation as a promising alternative for cancer immunotherapy. 相似文献
97.
Narita J Ishida S Okano K Kimura S Fukuda H Kondo A 《Applied microbiology and biotechnology》2006,73(2):366-373
The 5′-untranslated leader sequence (UTLS) of the slpA gene from Lactobacillus acidophilus contributes to mRNA stabilization by producing a 5′ stem and loop structure, and a high-level expression system for the lactic acid bacteria was developed using the UTLS in this study. A plasmid, which expresses α-amylase under the control of the ldh promoter, was constructed by integrating the core promoter sequence with the UTLS. The role of the UTLS in increasing the copies of the α-amylase mRNA was proved by measuring α-amylase activity in the culture supernatant and the relative expression of α-amylase mRNA was determined by the quantitative real-time PCR analysis. Moreover, several expression systems were constructed by combining the core promoter sequence with the UTLS or with the partially deleted UTLS and the expression level was evaluated. The use of the UTLS led to the success in improving α-amylase expression in the two strains of Lactobacillus casei and Lactococcus lactis. The current study showed that the improvement in protein production using the UTLS could be applied to the expression system in the lactic acid bacteria. 相似文献
98.
Revealing the control mechanisms responsible for the cell's surprisingly well-organized functions should lead directly to a better understanding of how the cell adapts to extraordinarily changing environments. A general framework for describing models that can represent diverse biochemical regulatory functions systematically would help not only systematic interpretation of the various models proposed for certain systems but also further understanding of the general control mechanism and design principles underlying different biological systems. This article presents a unified mathematical framework for describing gene regulatory units. The proposed framework is fairly compatible with the classical control theoretical framework, so it should serve as a connecting bridge between engineering control theory and biological control mechanisms. It should also provide a unified view of different regulatory units and facilitate systematic comparison of different mathematical models proposed in a variety of literature. 相似文献
99.
Regenerative therapies, including cell injection and bioengineered tissue transplantation, have the potential to treat severe
heart failure. Direct implantation of isolated skeletal myoblasts and bone-marrow-derived cells has already been clinically
performed and research on fabricating three-dimensional (3-D) cardiac grafts using tissue engineering technologies has also
now been initiated. In contrast to conventional scaffold-based methods, we have proposed cell sheet-based tissue engineering,
which involves stacking confluently cultured cell sheets to construct 3-D cell-dense tissues. Upon layering, individual cardiac
cell sheets integrate to form a single, continuous, cell-dense tissue that resembles native cardiac tissue. The transplantation
of layered cardiac cell sheets is able to repair damaged hearts. As the next step, we have attempted to promote neovascularization
within bioengineered myocardial tissues to overcome the longstanding limitations of engineered tissue thickness. Finally,
as a possible advanced therapy, we are now trying to fabricate functional myocardial tubes that may have a potential for circulatory
support. Cell sheet-based tissue engineering technologies therefore show an enormous promise as a novel approach in the field
of myocardial tissue engineering. 相似文献
100.
Harashima N Inao T Imamura R Okano S Suda T Harada M 《Cancer immunology, immunotherapy : CII》2012,61(5):667-676
Toll-like receptors (TLRs) are widely expressed in immune cells and play a crucial role in many aspects of the immune response.
Although some types of TLRs are also expressed in cancer cells, the effects and mechanisms of TLR signaling in cancer cells
have not yet been fully elucidated. In the present study, we analyzed the effects of polyinosinic-polycytidylic acid [poly(I:C)],
a TLR3 ligand, on three TLR3-expressing human prostate cancer cell lines (LNCaP, PC3, and DU145). We then further characterized
the underlying mechanisms, focusing on the poly(I:C)-sensitive LNCaP cell line. Poly(I:C) significantly reduced the viability
of LNCaP cells TLR3 and endosome dependently. One mechanism for the antitumor effect was caspase-dependent apoptosis, and
another mechanism was poly(I:C)-induced growth arrest. Cell survival and proliferation of LNCaP cells depended on the PI3K/Akt
pathway, and PI3K/Akt inhibitors induced apoptosis and growth arrest similar to poly(I:C) treatment. Additionally, poly(I:C)
treatment caused dephosphorylation of Akt in LNCaP cells, but transduction of the constitutively active form of Akt rendered
LNCaP cells resistant to poly(I:C). Immunoblot analysis of proliferation- and apoptosis-related molecules in poly(I:C)-treated
LNCaP cells revealed participation of cyclinD1, c-Myc, p53, and NOXA. Interestingly, poly(I:C) treatment of LNCaP cells was
accompanied by autophagy, which was cytoprotective toward poly(I:C)-induced apoptosis. Together, these findings indicate that
TLR3 signaling triggers apoptosis and growth arrest of LNCaP cells partially through inactivation of the PI3K/Akt pathway
and that treatment-associated autophagy plays a cytoprotective role. 相似文献