Search PubMed⌕ Search

Biomedical subjects

Satoshi Kaneko

Publications and source records attributed to Satoshi Kaneko.

At least 55 records · Page 3Linked to original sources

Site-directed incorporation of non-natural amino acid into Streptomyces xylanase.

A key residue, W26G existing around the substrate-binding cleft of beta-1,4-xylanase from Streptomyces olivaceoviridis E-86 (SoXyn10A), was ascertained in this study. Several kinds of W266 mutants substituted by Trp analogs were synthesized by a site-directed incorporation method to elucidate the functional role of this aromatic residue. Significant decreases of hydrolytic activities were observed in all mutants, even though the substitution of the aromatic side chain was not so drastic, suggesting that the W266 residue had a fatal role in the hydrolytic activity of SoXyn10A.

Amino Acids↗

Catalytic activity of tripeptidase from Lactococcus lactis to which amino acid substitution was introduced according to natural mutation.

Four mutations observed between tripeptidases from Lactococcus lactis subsp. lactis and subsp. cremoris were introduced one by one to the corresponding points in wild-type tripeptidase from L. lactis subsp. lactis. The k(cat) values of four resultant mutants were analyzed and discussed in stereographical terms. Change in catalytic activity appeared to be related to the sequential and steric location of mutation point within the enzyme protein, even though no drastic change was observed with one point mutation.

Amino Acid Sequence↗

[Analysis of facial lesions on adult type atopic dermatitis with anti-fungus drug (terbinafine hydrochloride) -- analysis of serum anti-Malassezia IgE antibody titers and histamine release test].

Malassezia furfur has been described as an aggravating factor in facial lesions of atopic dermatitis, and oral antifungal agents have been reported to be effective against these lesions. We used terbinafine hydrochloride to treat 15 patients with adult-type atopic dermatitis and evaluated its efficacy by measuring the improvement in facial skin manifestations, serum IgE values, and serum anti-Malassezia IgE antibody titers. A histamine release test (HRT) for Malassezia. was also performed in 6 of the 15 patients. The facial skin manifestations improved in 8 (53.3%) of the 15 patients, and there were significant simultaneous decreases in their serum IgE values. The serum anti-Malassezia IgE antibody titer decreased significantly in all 15 patients. However, no significant correlation was observed between the HRT and the facial skin manifestations. We concluded that oral terbinafine hydrochloride is effective against the facial lesions of atopic dermatitis patients and this is possibly caused by decrease of Malassezia antigen in the facial lesions.

Adult↗

Crystal structures of decorated xylooligosaccharides bound to a family 10 xylanase from Streptomyces olivaceoviridis E-86.

The family 10 xylanase from Streptomyces olivaceoviridis E-86 (SoXyn10A) consists of a GH10 catalytic domain, which is joined by a Gly/Pro-rich linker to a family 13 carbohydrate-binding module (CBM13) that interacts with xylan. To understand how GH10 xylanases and CBM13 recognize decorated xylans, the crystal structure of SoXyn10A was determined in complex with alpha-l-arabinofuranosyl- and 4-O-methyl-alpha-d-glucuronosyl-xylooligosaccharides. The bound sugars were observed in the subsites of the catalytic cleft and also in subdomains alpha and gamma of CBM13. The data reveal that the binding mode of the oligosaccharides in the active site of the catalytic domain is entirely consistent with the substrate specificity and, in conjunction with the accompanying paper, demonstrate that the accommodation of the side chains in decorated xylans is conserved in GH10 xylanases of SoXyn10A against arabinoglucuronoxylan. CBM13 was shown to bind xylose or xylooligosaccharides reversibly by using nonsymmetric sugars as the ligands. The independent multiple sites in CBM13 may increase the probability of substrate binding.

Binding Sites↗

Crystallization and preliminary X-ray crystallographic studies of alpha-galactosidase I from Mortierella vinacea.

alpha-Galactosidases catalyze the hydrolysis of a galactosyl residue from galactooligosaccharides and galactopolysaccharides. alpha-Galactosidase I from Mortierella vinacea was crystallized in two crystal forms using the hanging-drop vapour-diffusion method. Type 1 crystals belonged to space group I422, with unit-cell parameters a = b = 142.4, c = 131.5 A, and diffracted to beyond 2.1 A resolution, while type 2 crystals belonged to space group P4, with unit-cell parameters a = b = 100.9, c = 102.7 A, and diffracted to beyond 1.6 A resolution. This enzyme crystallized as a glycoprotein tetramer and the tetrameric structure was located around the crystallographic fourfold axis.

Bacterial Proteins↗

Akt phosphorylation and stabilization of X-linked inhibitor of apoptosis protein (XIAP).

Akt negatively regulates apoptotic pathways at a premitochondrial level through phosphorylation and modulation of proteins such as Bad, Forkhead proteins, and GSK-3beta. Akt has also been shown to protect cell death at a post-mitochondrial level, although its downstream targets have not been well documented. Here, we demonstrate that Akt, including AKT1 and AKT2, interacts with and phosphorylates X-linked inhibitor of apoptosis protein (XIAP) at residue serine-87 in vitro and in vivo. Phosphorylation of XIAP by Akt protects XIAP from ubiquitination and degradation in response to cisplatin. Moreover, autoubiquitination of XIAP is also inhibited by Akt. Consistent with this, an XIAP mutant introduced into cells which mimics the Akt-phosphorylated form (i.e. XIAP-S87D) displays reduced ubiquitination and degradation as compared with wild type XIAP. The greater stability of XIAP-S87D in cells translated to increased cell survival after cisplatin treatment. Conversely, a mutant that could not be phosphorylated by Akt (XIAP-S87A) was more rapidly degraded and showed increased cisplatin-induced apoptosis. Furthermore, suppression of XIAP by either siRNA or adenovirus of antisense of XIAP induced programmed cell death and inhibited Akt-stimulated cell survival in ovarian cancer cells. These data identify XIAP as a new downstream target of Akt and a potentially important mediator of the effect of Akt on cell survival.

Animals↗

Prospective study of screening for stomach cancer in Japan.

Although screening for stomach cancer is a widespread community service in Japan, the benefits of the screening program remain unclear. Our study investigated prospectively the relation between participation in stomach-cancer screening during the past 12 months and subsequent deaths. Data was derived from the Japan Collaborative Cohort Study, in which 480 stomach-cancer deaths were identified during an 8-year follow-up period. Cox proportional hazard regression was used to estimate the relative risk of death from stomach cancer and that from other causes while adjusting for potential confounding factors. In men, screening participation was associated significantly with a reduced risk of death from stomach cancer (relative risk [RR] = 0.54; 95% confidence interval [CI] = 0.41-0.70). The extent of the risk reduction was greater than potential health selection (for deaths other than stomach, RR = 0.71). The adjustment for potential confounding variables, however, attenuated the difference in risk of death (stomach cancer, RR = 0.65; other causes, RR = 0.71). In women, the magnitude of the association between screening participation and death from stomach cancer (RR = 0.74; 95% CI = 0.52-1.07) was equal to that for deaths from non-stomach cancers (RR = 0.74). Subgroup analysis showed that women with a parental history of stomach cancer had a reduced risk of death from stomach cancer associated with screening (RR = 0.32; 95% CI = 0.12-0.87). The present results underline the potential for selection bias in observational studies, and thus it remains an open question whether screening for stomach cancer is effective.

Adult↗

Crystal structure of rice alpha-galactosidase complexed with D-galactose.

alpha-Galactosidases catalyze the hydrolysis of alpha-1,6-linked galactosyl residues from galacto-oligosaccharides and polymeric galacto-(gluco)mannans. The crystal structure of rice alpha-galactosidase has been determined at 1.5A resolution using the multiple isomorphous replacement method. The structure consisted of a catalytic domain and a C-terminal domain and was essentially the same as that of alpha-N-acetylgalactosaminidase, which is the same member of glycosyl hydrolase family 27. The catalytic domain had a (beta/alpha)8-barrel structure, and the C-terminal domain was made up of eight beta-strands containing a Greek key motif. The structure was solved as a complex with d-galactose, providing a mode of substrate binding in detail. The d-galactose molecule was found bound in the active site pocket on the C-terminal side of the central beta-barrel of the catalytic domain. The d-galactose molecule consisted of a mixture of two anomers present in a ratio equal to their natural abundance. Structural comparisons of rice alpha-galactosidase with chicken alpha-N-acetylgalactosaminidase provided further understanding of the substrate recognition mechanism in these enzymes.

Amino Acid Sequence↗

Purification and characterization of alpha-galactosidase from sunflower seeds.

From 100 g sunflower seeds, 1.2 mg purified alpha-galactosidase was obtained with an overall yield of 51%. The alpha-galactosidase acted on both terminal alpha-galactosyl residues and side-chain alpha-galactosyl residues of the galactomanno-oligosaccharides and galactomannans. The cDNA coding for sunflower alpha-galactosidase was cloned and the deduced amino acid sequence revealed that the mature enzyme consisted of 363 amino acid residues with a molecular weight of 40,263. Seven cysteine residues were found but no putative N-glycosylation sites were present in the sequence. The deduced amino acid sequences of mature enzyme and alpha-galactosidases from coffee, guar and Mortierella vinacea alpha-galactosidase II showed over 81%, 77%, and 47% homology, respectively. These enzymes are classified into the third group in which the enzyme has no insertion sequence and a broad specificity on galactomanno-oligosaccharides compared to the other groups.

Amino Acid Sequence↗

Menstrual and reproductive factors and the mortality risk of gastric cancer in Japanese menopausal females.

OBJECTIVE: The relationship between gastric cancer and menstrual and reproductive history was investigated for 40,535 postmenopausal females from a large-scale prospective cohort in Japan (JACC: Japan Collaborative Cohort Study for Evaluation of Cancer Risk, sponsored by Monbusho). METHODS: A Cox proportional hazard model was used to estimate risk with respect to menstrual and reproductive factors for gastric cancer mortality. The effects on risk of potential confounders such as lifestyle, dietary habits, and socioeconomic status were controlled for using a stepwise procedure. RESULTS: During the study period (mean 8.2 years), 156 deaths due to gastric cancer were identified. Longer fertility tended to be inversely associated with risk of gastric cancer mortality, although the association was not significant. Late menopause had no obvious effect on gastric cancer risk. The risk ratio of delivery experience compared to no delivery was 0.62 (95% confidence interval 0.27-1.41), and this effect was consistent regardless of the number of deliveries. Early menarche was not associated with the risk of gastric cancer. The potential confounders were not seen to have distinct effects on the associations. CONCLUSIONS: Multiparity appears to confer a protective tendency on gastric cancer mortality; however, this result is inconsistent with previous reports. Further information needs to be accumulated from epidemiological studies in order to clarify the role of reproductive factors in gastric cancer.

Adult↗

Respiratory symptoms and cigarette smoking in 3,197 pulmonologist-based asthmatic patients with a highly prevalent use of inhaled corticosteroid.

The relation between smoking and risk of asthma has been well-examined; however little attention has been paid to the correlation between smoking and asthma symptoms. The aims of this study were to examine respiratory symptoms in asthmatics with a highly prevalent use of inhaled corticosteroid (ICS) and to assess the effects of smoking and its cessation. A cross-sectional study of pulmonologist-based 3197 asthmatics (men 45.2%, ages 20-97) was performed using a questionnaire about smoking habits, the incidence and frequency of symptoms (sputum, cough and wheezing, night symptoms, and shortness of breath), physical activity interference, and medication. Although 81.4% of the patients used ICS according to the international guideline, 14.9% had activity interference, and daily symptoms remained in 43.3%. At the time of the questionnaire, 21.6% were current and 25.1% were ex-smokers. In multiple logistic regression analysis, the factors of significance (p < 0.0001) were (1) smoking; for all four symptoms, (2) age and duration of asthma; for shortness of breath. Current smokers were at a risk of sputum (age-adjusted odds ratio 2.32 [95% confidence interval 1.73-3.11]; 2.09 [1.57-2.79]), of cough and wheezing (2.38 [1.81-3.14]; 1.78 [1.35-2.36]), of night symptoms (1.95 [1.41-2.60]; 1.47 [1.09-1.98]), and of shortness of breath (1.70 [1.26-2.28]; 1.30 [0.97-1.75]) in men and women, respectively. These ratios in ex-smokers decreased to the level similar to nonsmokers. Although 81.4% of asthmatic patients used ICS, 43.3% complained of daily respiratory symptoms, especially sputum. It is suggested that the effects of ICS on asthma symptoms may be interfered with by smoking and therefore more emphasis should be placed on cessation of smoking.

Administration, Inhalation↗

Leucyl/phenylalanyl (L/F)-tRNA-protein transferase-mediated N-terminal specific labelling of a protein in vitro.

N-terminal specific radioisotope (RI) labelling of a protein posessing lysine as the N-terminal residue (Lys-XBD) was achieved under mild enzymatic reaction condition using L/F-tRNA-protein transferase. To expose a single lysine moiety only at the N-teminal site, we overexpressed pelB signal peptide-Lys-XBD fusion protein using the bacterial expression host BL21(DE3). The pelB signal peptide was sponteneously cleaved in E. coli to obtain the desired Lys-XBD.

Amino Acid Sequence↗

Projection of lung cancer mortality in Japan.

According to the National Vital Statistics data, age-standardized mortality rates (ASRs) of lung cancer have shown slightly declining trends in Japan for both men and women. In order to evaluate whether this tendency will continue, a Bayesian age-period-cohort (APC) model was applied using the National Vital Statistics data from 1952 to 2001. In the projection, a Gaussian autoregressive prior model was applied to smooth age, period, and cohort effects from its 2 immediate predecessors by extrapolation. Posterior distributions from which we drew inferences on mortality rates were derived from 15,000 iterations using 5000 burn-in iterations. We defined the median of the iterated values as the overall summary mortality rate of the iterated results. Our results suggest that the number of deaths due to lung cancer will double for men and women during the next 3 decades due to the aging of the baby-boomer generation (individuals who were born between 1947 and 1951). Currently declining trends in some age groups will reverse and start to increase again in the next decades. However, for recent birth cohorts, the results of the projection varied according to whether the data set included early age group mortality or not. Lung cancer mortality in the future depends on the risk factors engaged in by today's young people, especially smoking. Strong promotion of anti-smoking measures and careful surveillance for lung cancer are needed.

Adult↗

Purification and characterization of extracellular beta-galactosidase secreted by supension cultured rice (Oryza sativa L.) cells.

A beta-galactosidase was purified 1300-fold by lactosyl-Sepharose 4B and Sephacryl S-200 column chromatographies from the cultured medium of a rice-cell suspension. The purified enzyme appeared as 47 kD and 40 kD polypeptides on SDS-PAGE and had a specific activity of 65.1 units/mg. Optimum activity was observed at pH 3.5 and 60 degrees C. The enzyme released galactose from galactoxyloglucan and pectic galactans.

Chromatography, Liquid↗

Neurokinin-1 and neurokinin-2 antagonism inhibits long-term acid fog-induced airway hyperresponsiveness.

BACKGROUND: We recently reported that airway hyperresponsiveness (AHR) induced by a 6-h exposure to sulfuric acid (H(2)SO(4)) was inhibited by either the neurokinin (NK)-1 receptor antagonist, FK888, or the NK-2 receptor antagonist, SR48968, when administered immediately before the exposure. The aims of this study were to determine whether these antagonists have any therapeutic efficiency against AHR after long-term H(2)SO(4) inhalation and to elucidate the mechanisms in ovalbumin sensitized guinea pigs. METHODS: Specific airway resistance (sRaw), AHR, and BAL fluid were analyzed after an 8-week exposure to H(2)SO(4) aerosol (82 mg/m(3), pH 1.7, 40 mOsm) or hypotonic saline solution (pH 5.9, 40 mOsm) as a control. The H(2)SO(4) group then received a 2-week treatment with FK888, SR48968, or vehicle. RESULTS: The AHR and the eosinophil count in BAL fluid were significantly increased in the H(2)SO(4) group compared to control animals, while sRaw was significantly elevated in both groups after the 8-week exposure. Treatment with both FK888 and SR48968 significantly reduced the AHR and tended to inhibit eosinophilia in BAL fluid, but sRaw did not change. The degree of AHR improvement with SR48968 was much larger than with FK888. CONCLUSION: Our results show that both NK-1 and NK-2 receptor antagonists inhibited long-term H(2)SO(4)-induced AHR in sensitized guinea pigs, and the effect was much greater with an NK-2 antagonist. We suggest that NK-1 or NK-2 antagonism might partially inhibit the H(2)SO(4)-induced influx of eosinophils into the lung.

Airway Resistance↗