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A rapid and sensitive assay for [3H]GTP binding activity of tubulin has been developed. This assay method is based on the quantitative retention of [3H]GTP. Tubulin complex on a nitrocellulose membrane filter. It was also found that bovine brain tubulin is markedly stablized by glycerol and GTP against denaturation. A large-scale purification of bovine brain tubulin was achieved using the new assay procedure and by the inclusion of glycerol and GTP in a buffer solution used for column chromatograph. The purified tubulin could be stored at -80degrees in the presence of glycerol and GTP for at least a year without any apprecialbe loss of [3H]GTP- and [3H]colchicine binding activities. The interaction of tubulin with guanine nucleotides was also studied using the nitorcellulose membrane filter procedure. It was found that the binding of [3H]GTP to tubulin with an empty exchangeable site proceeded promptly within k sec while the exchange of [3H]GTP- with a GTP-tubulin complex in which the exchangeable site had been occupied with unlabeled GTP occured more slowly. The dissociation constants for GTP and GDP at the exchangeable site of tubulin were determined as 0.5 times 10-6M and 1.9 times 10-6M, respectively. 5'-Guanylylimidodiphosphate could interact, although less strongly, with tubulin at this site, whereas the interaction of other nucleoside triphosphates includint ATP, CTP, UTP, and 5'-guanylyl methylenediphosphonate was very weak, if it occured at all. The presence of Mg2+ and a free sulfhydryl group was found to be essential for binding of [3H]GTP to tubulin. Ca2+ was found to replace Mg2+ in this binding reaction. 相似文献
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Kyoko Saio Emiko Koyama Tokuji Watanabe 《Bioscience, biotechnology, and biochemistry》2013,77(10):1195-1200
This paper constitutes the first report in a research series undertaken in order to clarify the interactive relationships among soybean meal protein, calcium and phytic acid in solution and in precipitate.Data presented are mainly concerned with solubility characteristics of soybean meal proteins as a function of pH. The results support the suggestion as follows; phosphorus and calcium give the remarkable effects on solubility characteristics of protein, the complex containing protein, calcium and phosphorus is formed above isoelectric point of protein and the combinations are labile above at pH 8, especially by heating. 相似文献
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Suzuki Setsuko Kyoko Sugai Ichiro Tamaki Koji Takayama Hidetoshi Kato Hiroshi Yoshimaru 《Molecular ecology》2020,29(6):1050-1068
Pandanus boninensis, endemic to the Ogasawara Islands, Japan, is distributed on both the older Bonin and younger Volcano Islands. In this study, we conducted population genetic analyses of P. boninensis on these islands to examine the population diversity and structure across old and young islands, to assess potential differences in population demography with island age, and to collect any evidence of migration between old and young islands. We found that the genetic diversity of expressed sequence tag (EST)–based microsatellite (SSR) markers, the nucleotide diversity of nuclear DNA sequences, and the haplotype diversity of chloroplast DNA on young islands were lower than those on old islands. Clustering analyses of EST‐SSR indicated that populations on old islands were strongly diverged from those on young islands. Approximate Bayesian computation analysis of EST‐SSR suggested that population expansion occurred on old islands while population reduction occurred on young islands. We also found evidence of migration among old islands (mostly from south to north), while it appears that there have been very few migration events between old and young islands. These differences could be due to the fact that young islands tend to be geographically isolated and support smaller populations that began a shorter time ago from limited founders. The P. boninensis populations on the Volcano Islands are interesting from an evolutionary perspective as they constitute a classic example of the early stages of progressive colonization on oceanic islands with small effective population sizes and low genetic diversity. 相似文献
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Aida Rieko Hagiwara Keitaro Okano Kazunori Nakata Kyoko Obata Yuho Yamashita Takahiro Yoshida Kaoru Hagiwara Hiromi 《Molecular biology reports》2021,48(3):2291-2297
Molecular Biology Reports - Apigenin is a flavonoid with antioxidant and anticancer effects. It has been reported that apigenin inhibits proliferation, migration, and invasion and induces apoptosis... 相似文献
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Kume Manabu Nakayama Naoki Iwasaki Yasunari Hori Tomoya Watanabe Shun Terashima Yuki Medo Ayano Arai Nobuaki Yamashita Yoh Mitamura Hiromichi 《Environmental Biology of Fishes》2021,104(4):529-533
Environmental Biology of Fishes - Eel movement patterns have been frequently studied to learn about their movements within the fresh- and brackish waters of the same river before their spawning... 相似文献