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Molecular characterization of variant alpha-subunit of electron transfer flavoprotein in three patients with glutaric acidemia type II--and identification of glycine substitution for valine-157 in the sequence of the precursor, producing an unstable mature protein in a patient. 总被引:1,自引:0,他引:1 下载免费PDF全文
In our previous study of eight glutaric acidemia type II (GAII) fibroblast lines by using [35S]methionine labeling and immunoprecipitation, three of them had a defect in the synthesis of the alpha-subunit of electron transfer flavoprotein (alpha-ETF) (Ikeda et al. 1986). In one of them (YH1313) the labeling of the mature alpha-ETF was barely detectable, while that of the precursor (p) was stronger. In another (YH605) no synthesis of immunoreactive p alpha-ETF was detectable. In the third cell line (YH1391) the rate of variant p alpha-ETF synthesis was comparable to normal, but its electrophoretic mobility was slightly faster than normal. In the present study, the northern blot analysis revealed that all three mutant cell lines contained p alpha-ETF mRNA and that their size and amount were comparable to normal. In immunoblot analysis, both alpha- and beta-ETF bands were barely detectable in YH1313 and YH605 but were detectable in YH1391 in amounts comparable to normal. Sequencing of YH1313 p alpha-ETF cDNA via PCR identified a transversion of T-470 to G. We then devised a simple PCR method for the 119-bp section (T-443/G-561) for detecting this mutation. In the upstream primer, A-466 was artificially replaced with C, to introduce a BstNI site into the amplified copies in the presence of G-470 from the variant sequence. The genomic DNA analysis using this method demonstrated that YH1313 was homozygous for T----G-470 transversion. It was not detected either in two other alpha-ETF-deficient GAII or in seven control cell lines. The alpha-ETF cDNA sequence in YH605 was identical to normal. 相似文献
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Kinoshita Kodzue Indo Yoriko Tajima Tomoyuki Kuze Noko Miyakawa Etsuko Kobayashi Toshio Nakamura Tomoyuki Ogata Mitsuaki Okumura Fumihiko Hayakawa Takashi Morimura Naruki Mori Yusuke Okamoto Munehiro Ozaki Yasuhiko Hirata Satoshi 《Primates; journal of primatology》2021,62(3):475-475
Primates - In the original publication of the article, the coauthor “Takashi Hayakawa” was wrongly assigned as co-corresponding author. 相似文献
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Toshio Yoshida Akira Komatsu Motoichi Indo 《Bioscience, biotechnology, and biochemistry》2013,77(6):433-437
Menthols were converted to Δ3-menthone enol acetate (VII) via menthones having only one asymmetric carbon atom. It was shown that the optical rotation of menthone enol acetate was proportional to the optical purity of starting menthols. Optical purity of original menthol could, therefore, be determined by optical rotation of menthone enol acetate derived from. 相似文献
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Evidence of ROS generation by mitochondria in cells with impaired electron transport chain and mitochondrial DNA damage 总被引:6,自引:0,他引:6
Indo HP Davidson M Yen HC Suenaga S Tomita K Nishii T Higuchi M Koga Y Ozawa T Majima HJ 《Mitochondrion》2007,7(1-2):106-118
Mitochondrial damage is a well known cause of mitochondria-related diseases. A major mechanism underlying the development of mitochondria-related diseases is thought to be an increase in intracellular oxidative stress produced by impairment of the mitochondrial electron transport chain (ETC). However, clear evidence of intracellular free radical generation has not been clearly provided for mitochondrial DNA (mtDNA)-damaged cells. In this study, using the novel fluorescence dye, 2-[6-(4'-hydroxy)phenoxy-3H-xanthen-3-on-9-yl]benzoic acid (HPF), which was designed to detect hydroxyl radicals (*OH), intracellular free radical formation was examined in 143B cells (parental cells), 143B-rho(0) cells (mtDNA-lacking cells), 87 wt (cybrid), and cybrids of 4977-bp mtDNA deletion (common deletion) cells containing the deletion with 0%, 5%, 50% and >99% frequency (HeLacot, BH5, BH50 and BH3.12, respectively), using a laser confocal microscope detection method. ETC inhibitors (rotenone, 3-nitropropionic acid, thenoyltrifluoroacetone, antimycin A and sodium cyanide) were also tested to determine whether inhibitor treatment increased intracellular reactive oxygen species (ROS) generation. A significant increase in ROS for 143B-rho(0) cells was observed compared with 143B cells. However, for the 87 wt cybrid, no increase was observed. An increase was also observed in the mtDNA-deleted cells BH50 and BH3.12. The ETC inhibitors increased intracellular ROS in both 143B and 143B-rho(0) cells. Furthermore, in every fluorescence image, the fluorescence dye appeared localized around the nuclei. To clarify the localization, we double-stained cells with the dye and MitoTracker Red. The resulting fluorescence was consistently located in mitochondria. Furthermore, manganese superoxide dismutase (MnSOD) cDNA-transfected cells had decreased ROS. These results suggest that more ROS are generated from mitochondria in ETC-inhibited and mtDNA-damaged cells, which have impaired ETC. 相似文献
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New double stimuli-responsive poly(alpha-N-substituted gamma-glutamine) has been developed, which was synthesized by the reaction of poly(gamma-glutamic acid) with amino alcohols. Appropriate combinations of the amino alcohols provided the biodegradable poly(amino acid) exhibiting a sharp lower critical solution temperature (LCST) in water. Furthermore, the phase transition temperature was highly sensitive to pH changes. 相似文献
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Toshio Yoshida Akira Komatsu Motoichi Indo 《Bioscience, biotechnology, and biochemistry》2013,77(9):824-831
The isomerization and racemization of menthol isomers have been investigated with copper chromite and Raney nickel catalyst or sodium mentholates. The equilibrium concentration has been found to correspond to 55~56% of menthol with the catalytic isomerization, whereas to 74~75% of menthol with sodium mentholates. From these data, the free energy difference between equatorial and axial group in menthols is calculated to be about 0.5~0.6 Kcal./mole for hydroxyl and 1.4~1.5 Kcal./mole for methyl at 200°C. 相似文献
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Toshio Yoshida Akira Komatsu Motoichi Indo 《Bioscience, biotechnology, and biochemistry》2013,77(1):68-72
The hitherto unknown cyanoacetates of menthol isomers, i.e., (±)-menthyl, (±)- and (+)-neomenthyl, and (±)- and (+)-isomenthyl cyanoacetate, have been prepared. It has been proved to be possible to isolate effectively pure (±)-menthol from optically inactive mixture of isomers through the cyanoacetate, followed by saponification. 相似文献
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Toshifumi Fukuda Yoshikichi Higuchi Motoichi Matsuda 《Bioscience, biotechnology, and biochemistry》2013,77(5):417-420
Authors succeeded in rearing eri-silkworms from hatching to the mouth stage on an artificial food. The artificial food devised were composed of powdered leaves of castor-oil plant, “Kinako” (a powder of parched soy bean), sucrose, agar-agar, inhibitor solution and water. The results on larval raising, cocoon fibres and eggs of the eri-silkworms reared on artificial food were the same as those in the case of the eri-silkworms reared on fresh leaves of castor-oil plant, natural food for this insect, or rather better. The raising of eri-silkworms on artificial food may be adopted industrially for production of a wild silk. 相似文献