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Kazuyuki Morihara Tatsushi Oka Hiroshige Tsuzuki Yoshiharu Tochino Takashi Kanaya 《Biochemical and biophysical research communications》1980,92(2):396-402
We have established a procedure for converting porcine insulin into human insulin using a serine protease from M497-1 which shows unique specificity against lysine residues on the carboxyl side of the splitting point. Desalanine-(B30)-insulin (DAI) was prepared by digestion of porcine insulin with protease. The coupling between DAI and Thr-OBut was performed by the same enzyme at pH 6.5 with a large excess of the amine component (Thr-OBut) in the presence of high concentrations of organic co-solvents. The highest yield was 85% by 20 h reaction at 37°C. The synthesized [Thr-OBut-B30]-insulin was isolated, then deprotected with trifluoroacetic acid in the presence of anisole to obtain semisynthetic human insulin. 相似文献
3.
Jun Ishizaki Mikio Tamaki Masaru Shin Hiroshige Tsuzuki Kazumasa Yoshikawa Hiroshi Teraoka Nobuo Yoshida 《Applied microbiology and biotechnology》1992,36(4):483-486
Summary Recombinant human glucagon was succesfully produced with a high level of expression in Escherichia coli as a fusion protein with human interferon . The synthetic gene was designed to release glucagon, which does not contain glutamic acid residues, from fusion protein with the Staphylococcus aureus strain V8 protease that specifically cleaves the peptide bond on the carboxyl side of the glutamic acid residue. The resulting glucagon was purified to homogeneity by a combination of C18 reverse-phase HPLC and ion-exchange HPLC. The yield of intact glucagon obtained from 11 of culture was approximately 12 mg. The structure of recombinant human glucagon was confirmed by HPLC and amino acid composition/sequence analyses.
Offprint requests to: J. Ishizaki 相似文献
4.
Mouse fibroblasts, 3T3 cells, require a solid surface for continuous growth, but when 3T3 cells, during their exponential phase in Petri dishes, were transferred to a suspension culture, the number of cells roughly doubled by 30 h. During the suspension culture the number of pairing cells (c2) increased, but that of the single cells decreased. When cells synchronized at mitosis or at the G1-S boundary were transferred to the suspension culture, the number of pairing cells peaked at 30 min and at 10 h, respectively. DNA synthesis began immediately after the cells, which were cultured for 16 h in the suspension, had settled onto the surface of the Petri dishes. When cells in a confluent culture were arrested at an early G1 period and were suspended, the number of pairing cells did not increase. These results indicate that the most important locus for anchorage growth seems to be at a late G1 period of the cell cycle. 相似文献
5.
A winter geometrid moth, Inurois punctigera, shows sympatric and genetically isolated seasonal populations (i.e. early‐ and late‐winter populations) in the cold regions of Japan, whereas it shows only mid‐winter populations in the warm regions. Variation in adult flight phenology on a large geographic scale along latitudinal environmental gradients has been described, but the phenological variation on a more local scale along altitudinal environmental gradients has not yet been characterized. In the present study, we assessed the flight phenology at high‐ and low‐elevation areas in Mt. Rokko, Hyogo, Japan. First, we revealed that flight period was not disrupted in mid‐winter, even at high‐elevation areas (>660 m) but the population abundance was much lower in high‐elevation areas than in low‐elevation areas. Then, in the following two seasons, we investigated I. punctigera abundance, winter harshness (i.e. winter temperature) and their host plant abundance in nine closely located stations in Mt. Rokko. A generalized linear mixed model analysis indicated a greater effect of winter temperature on I. punctigera abundance compared to available food resources, suggesting that differences in winter harshness among elevation shapes the gradient of I. punctigera abundance along altitude. Our findings suggest that harsh conditions during winter function as selective agents on mid‐winter types of I. punctigera, and this could be involved in the divergence between sympatric early‐ and late‐winter populations of I. punctigera. 相似文献
6.
Yuichi Takashi Shun Sawatsubashi Itsuro Endo Yukiyo Ohnishi Masahiro Abe Munehide Matsuhisa Daiji Kawanami Toshio Matsumoto Seiji Fukumoto 《Biochemistry and Biophysics Reports》2021
Fibroblast growth factor (FGF) 23 produced by the bone is the principal hormone to regulate serum phosphate level. Serum FGF23 needs to be tightly regulated to maintain serum phosphate in a narrow range. Thus, we hypothesized that the bone has some phosphate-sensing mechanism to regulate the production of FGF23. Previously we showed that extracellular phosphate induces the phosphorylation of FGF receptor 1 (FGFR1) and FGFR1 signaling regulates the expression of Galnt3, whose product works to increase FGF23 production in vitro. In this study, we show the significance of FGFR1 in the regulated FGF23 production and serum phosphate level in vivo. We generated late-osteoblast/osteocyte-specific Fgfr1-knockout mice (Fgfr1fl/fl; OcnCre/+) by crossing the Ocn-Cre and the floxed Fgfr1 mouse lines. We evaluated serum phosphate and FGF23 levels, the expression of Galnt3 in the bone, the body weight and life span. A selective ablation of Fgfr1 aborted the increase of serum active full-length FGF23 and the enhanced expression of Galnt3 in the bone by a high phosphate diet. These mice showed more pronounced hyperphosphatemia compared with control mice. In addition, these mice fed with a control diet showed body weight loss after 23 weeks of age and shorter life span. These results reveal a novel significance of FGFR1 signaling in the phosphate metabolism and normal life span. 相似文献
7.
Miyata Seiko Otake Hironao Fujishiro Hiroshige Iwamoto Kunihiro Noda Akiko Sone Michihiko Ozaki Norio 《Sleep and biological rhythms》2022,20(3):403-411
Sleep and Biological Rhythms - Sleep problems and obstructive sleep apnea (OSA) increase with age and disturb life in old age. Positional therapy is one option to treat OSA, but the differences in... 相似文献
8.
Balb/c 3T3 cells have long been used in cell biological studies. When cells were treated with 2 mM sodium butyrate they became flattened and their nuclei became small relative to the cytoplasm. Electron micrographically, intracellular organellae developed significantly: the number of mitochondria increased and membranous structures, namely the rough-surfaced endoplasmic reticulum and Golgi apparatus, developed markedly. Blood coagulation factor VIII was induced in the presence of 2 mM sodium butyrate in a cell sonicate, but not in culture medium. The amount of this factor was almost 100-fold that in the nontreated cells 4 days after culture initiation, indicating that the cloned Balb/c 3T3 cells were of endothelial origin. 相似文献
9.
Tsuji S Uehori J Matsumoto M Suzuki Y Matsuhisa A Toyoshima K Seya T 《The Journal of biological chemistry》2001,276(26):23456-23463
Galactofuranosyl residues are present in various microorganisms but not in mammals. In this study, we identified a human lectin binding to galactofuranosyl residues and named this protein human intelectin (hIntL). The mature hIntL was a secretory glycoprotein consisting of 295 amino acids and N-linked oligosaccharides, and its basic structural unit was a 120-kDa homotrimer in which 40-kDa polypeptides were bridged by disulfide bonds. The hIntL gene was split into 8 exons on chromosome 1q21.3, and hIntL mRNA was expressed in the heart, small intestine, colon, and thymus. hIntL showed high levels of homology with mouse intelectin, Xenopus laevis cortical granule lectin/oocyte lectin, lamprey serum lectin, and ascidian galactose-specific lectin. These homologues commonly contained no carbohydrate recognition domain, which is a characteristic of C-type lectins, although some of them have been reported as Ca(2+)-dependent lectins. Recombinant hIntL revealed affinities to d-pentoses and a d-galactofuranosyl residue in the presence of Ca(2+), and recognized the bacterial arabinogalactan of Nocardia containing d-galactofuranosyl residues. These results suggested that hIntL is a new type lectin recognizing galactofuranose, and that hIntL plays a role in the recognition of bacteria-specific components in the host. 相似文献
10.