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261.
Formation of free radicals in golden hamster embryo (GHE) cells in the frozen living state by gamma irradiation has been studied by electron spin resonance spectroscopy at 4.2 and 77 K. The relative yields of H atoms, OH radicals, and organic radicals trapped in the irradiated GHE cells are 12, 72, and 16%, respectively, of total radical yields. When dimethylsulfoxide (DMSO) is added to GHE cells at 77 K, a large quantity of CH2SOCH3 radicals (DMSO radicals) are formed after gamma irradiation. The yields of OH radicals are not affected by the addition of DMSO. When the GHE cell-DMSO mixtures are irradiated with gamma rays at 77 K and then warmed to 111 K, the OH radicals decay, whereas the DMSO radicals do not increase complementarily. Moreover, the decay rates of the OH radicals at 111 K do not depend upon the concentration of DMSO. Thus OH radicals do not react with DMSO during warming of the irradiated sample. When H atoms are produced by gamma irradiation of acid ice at 60 K, the decay rates of the H atoms at 77 K increase with increasing DMSO concentration, indicating that DMSO reacts with H atoms (CH3SOCH3 + H----.CH2SOCH3 + H2) at 77 K by quantum-mechanical tunneling. When the GHE cell-DMSO mixture is irradiated with gamma rays at 77 or 4.2 K in the dark, DMSO ions are produced in addition to DMSO radicals. Therefore it is concluded that DMSO does not scavenge OH radicals, but does capture H atoms, holes and/or electrons in the gamma-irradiated cells, resulting in the remarkable formation of DMSO radicals. This scavenger effect of DMSO may be related to the radioprotection of DMSO against cell killing described in the companion paper (Watanabe et al., Radiat. Res., this issue). 相似文献
262.
A protein kinase C alpha (PKC alpha) cDNA confers increased phorbol ester binding activity to intact cells when transiently expressed in COS cells or expressed stably in transfected rat 3Y1 fibroblasts. A point mutant (PKC alpha K----R) of PKC alpha, where Lys368 at the putative ATP-binding site is replaced with Arg, confers enhanced phorbol ester binding activity to both transiently and stably expressed COS and 3Y1 cells, respectively. Like endogenous and exogenously expressed wild type PKC alpha, the mutant PKC alpha K----R is translocated from the cytosol to the particulate fraction when cells are treated with a phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA). On the other hand, the mutant PKC alpha K----R is not degraded when cells are treated with TPA, making a clear contrast to wild type PKC alpha; i.e. the mutant is resistant to TPA-mediated down-regulation. The mutant lacks kinase activity as expected, as judged by autophosphorylation and by a kinase assay using a peptide substrate, although the phorbol ester binding activity remains intact. These results suggest a link between the kinase activity of PKC alpha and the sensitivity to TPA-mediated proteolytic degradation. We propose that autophosphorylation of PKC alpha is a prerequisite for proteolytic cleavage associated with the down-regulation of PKC alpha. 相似文献
263.
Saburo Tamura Ching-Fun Chang Akinori Suzuki Sumio Kumai 《Bioscience, biotechnology, and biochemistry》2013,77(3):391-397
Two new isoflavonoids were isolated from red clover as germination inhibitors for the same plant and their structures were determined as a glucoside of biochanin A (7-d-β-glucosyl-5,7-dihydroxy-4′-methoxyisoflavone) (II) and its 5-malonate (I), respectively. Besides these compounds the following substances were also isolated as inhibitors: trifolirhizin (III), ononin (IV), daidzein (V) and its 7-glucoside (VI), formononetin (VII), genistein (VIII) and biochanin A (IX). 相似文献
264.
Akihiro Okitani Atsushi Suzuki Ryung Yang Masao Fujimaki 《Bioscience, biotechnology, and biochemistry》2013,77(12):2135-2141
The effect of cathepsin D and pepsin treatment on rabbit myofibril was studied by measuring the amount of proteolytic products and Mg-enhanced ATPase activity.When myofibril was treated with cathepsin D at 3°C and pH 5.0 or 5.5, a little but detectable amount of nonprotein nitrogenous compounds was released. However, there was no change in ATPase activity of myofibril, though treated with cathepsin D of higher units than assumed to be in muscle.When myofibril was treated with pepsin under the same condition as used above, there was an increase in KCl-concentration dependence of ATPase activity followed by a decrease in the maximal value of ATPase activity.From the present results, it was concluded that cathepsin D might not take a main role on the post-mortem degradation of myofibril. 相似文献
265.
Yoichiro Fujioka Shinya Nishide Toyoyuki Ose Tadaki Suzuki Izumi Kato Hideo Fukuhara Mari Fujioka Kosui Horiuchi Aya O. Satoh Prabha Nepal Sayaka Kashiwagi Jing Wang Mika Horiguchi Yuko Sato Sarad Paudel Asuka Nanbo Tadaaki Miyazaki Hideki Hasegawa Yusuke Ohba 《Cell host & microbe》2018,23(6):809-818.e5
266.
Photosynthetically competent chloroplasts were isolated fromcells of Euglena gracilis Z grown photoautotrophically in 1.5%CO2. The isolated chloroplasts were intact and substantiallyfree from cytosolic, mitochondrial and microbody materials.The effects of some compounds on the activity of photosynthetic14CO2 fixation were examined. The optimal pH and sorbitol concentrationwere 8.0 and 0.33 M, respectively. The chloroplasts requireda high level of P, (5 to 20 mM) for the maximal rate of photosynthesis.They were insusceptible to 10 mM of free Mg2+. ATP, ADP andAMP at 1 to 5 mM notably stimulated photosynthesis, althoughhigh concentrations of AMP were unfavorable. In the assay mediumdeveloped for this study, the chloroplasts exhibited photosyntheticactivity of 120µmoles-mg1 Chl-h1 at 30?C. Chloroplasts could also be isolated from cells grown under ordinaryair. The rate of photosynthetic 14CO2 fixation at 1 mM NaHl4CO3was higher in these chloroplasts than in those isolated fromcells grown in 1.5% CO2, whereas at 10 mM NaHl4CO3, the ratesof the two types of chloroplasts were nearly the same. Theseresults suggest that the CO2 concentration given during growthof the algal cells affects the affinity for dissolved inorganiccarbon at the chloroplast level. (Received March 30, 1987; Accepted August 17, 1987) 相似文献
267.
H Suzuki M Obara H Kuwayama T Kanazawa 《The Journal of biological chemistry》1987,262(32):15448-15456
Sarcoplasmic reticulum vesicles were modified with a fluorescent thiol reagent, N-iodoacetyl-N'-(5-sulfo-1-naphthyl)ethylenediamine. One mol of readily reactive thiols per mol of the Ca2+-ATPase was labeled without a loss of the catalytic activity. The fluorescence of the label increased by 8% upon binding of Ca2+ to the high affinity sites of the enzyme. This fluorescence enhancement probably reflects a conformational change responsible for Ca2+-induced enzyme activation. Upon addition of ATP to the Ca2+-activated enzyme, the fluorescence decreased by 15%. This fluorescence drop and formation of the phosphoenzyme intermediate were determined under the same conditions with a stopped-flow apparatus and a rapid quenching system. The amplitude of the fluorescence drop thus determined was saturated with 3 microM ATP. This shows that the fluorescence drop was caused by ATP binding to the catalytic site. In contrast, the rate of the fluorescence drop was not saturated even with 50 microM ATP. The fluorescence drop coincided with phosphoenzyme formation at 0.5 or 3 microM ATP, but it became much faster than phosphoenzyme formation when the ATP concentration was raised to 100 microM. These results indicate that the ATP-induced fluorescence drop reflects a conformational change in the enzyme.ATP complex. The fluorescence drop was accompanied by a red spectrum shift, which suggests that the label was exposed to a more hydrophilic environment. The electrophoretic analysis of the tryptic digest of the labeled enzyme (10.9 kDa) showed that almost all of the label was located on the 5.2-kDa fragment which includes the carboxyl terminus and the putative ATP-binding domain. The sequencing of the two major labeled peptides, which were isolated from the thermolytic digest of the labeled enzyme, revealed that the labeled site in either of these peptides was Cys674. It seems likely that the label bound to this Cys674 could be involved in the observed fluorescence changes. 相似文献
268.
269.
Tomohiko Suzuki 《The protein journal》1994,13(1):9-13
The cDNA for the unusual 41 kD myoglobin of the abaloneNordotis madaka was amplified by polymerase chain reaction (PCR), and the cDNA-derived amino acid sequence of 378 residues was determined. As with the myoglobin of the related abaloneSulculus diversicolor (Suzuki and Takagi,J. Mol. Biol. 228, 698–700, 1992), the sequence ofNordotis myoglobin showed no significant homology with any other globins, but showed high homology (35% identity) with vertebrate indoleamine 2,3-dioxygenase, a tryptophan degrading enzyme containing heme. The amino acid sequence homology betweenNordotis andSulculus myoglobins was 87%. These results support our previous idea that the abalone myoglobins evolved from a gene for indoleamine dioxygenase, but not from a globin gene, and therefore all of the hemoglobins and myoglobins are not homologous. Thus, abalone myoglobins appear to be a typical case of convergent evolution. 相似文献
270.
Anthony O'L Richards Stephen H. Stanley Motoshi Suzuki Howard Dalton 《Biocatalysis and Biotransformation》1994,8(4):253-267
Methylococcus capsulatus (Bath) possesses methane monooxygenases (soluble - (sMMO) and particulate - (pMMO)) which are able to catalyse the epoxidation of propylene to propylene oxide. In a previous paper we have shown that the production of the epoxide caused a rapid inactivation of the bioconversion process (Stanley et al, 1992). This paper shows that cultures containing pMMO, inactivated by propylene oxide production, could be completely reactivated in the presence of growth substrates within 5 h after the removal of propylene oxide so long as the propylene oxide production rate was below 150 nmol min-1 [mg dry weight cells]-1. Reactivation under these conditions was detectable within 30 min of propylene oxide removal. On the other hand, cells inactivated by propylene oxide production rates in excess of 150 nmol min-1 [mg dry weight]-1 did not begin to recover activity within the 30 min period. Furthermore a lag period was observed before reactivation began which was dependent upon the initial production rate. Cultures possessing sMMO took twice as long to recover their activity compared with cells containing pMMO.
Reactivation of propylene oxide production could occur without growth, but the process required the presence of a carbon and energy source (methane or methanol), sulphur, nitrogen and oxygen, although copper (which is normally involved in pMMO activity) was not required. It was shown that de novo protein synthesis was required for reactivation of activity.
Production rates of 12 g 1-1 d-1 could be maintained for longer than three weeks in a single phase production process and rates up to 30 g 1-1 d-1 were achieved in a two stage process. Using Methylocystis parvus (OBBP) rates of up to 90 g 1-1 d-1 were attained over a one week period. 相似文献
Reactivation of propylene oxide production could occur without growth, but the process required the presence of a carbon and energy source (methane or methanol), sulphur, nitrogen and oxygen, although copper (which is normally involved in pMMO activity) was not required. It was shown that de novo protein synthesis was required for reactivation of activity.
Production rates of 12 g 1-1 d-1 could be maintained for longer than three weeks in a single phase production process and rates up to 30 g 1-1 d-1 were achieved in a two stage process. Using Methylocystis parvus (OBBP) rates of up to 90 g 1-1 d-1 were attained over a one week period. 相似文献