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141.
Tetsuya Tanigawa Yasuo Mizo-oku Kouichi Moriguchi Takashi Suzuki Takahiko Osumi Masaaki Odomi 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1996,683(2):135
A simple and rapid quantitative method for 13C-labelled urea ([13C]urea) in human serum was developed by using high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (HPLC-APCI-MS). This method is used to establish and normalize the [13C]urea breath test, which is considered as an effective diagnostic method for Helicobacter pylori infection. HPLC-APCI-MS, involving a simple pretreatment process such as diluting serum with water, was shown to be able to discriminate the extrinsic [13C]urea from intrinsic urea present at high concentration in serum. In addition, a 13C nuclear magnetic resonance spectroscopic quantitative method for [13C]urea in human urine is also described. The precision and accuracy of measured concentrations in these two methods were found to be within the acceptable limit. An application of these methods to investigate the pharmacokinetic profile of orally administered [13C]urea in human serum and urine is also presented. 相似文献
142.
Shinichi Yoshida Saori Yonehara Shigemi Minami Hyo-cheol Ha Kenji Iwahara Takashi Watanabe Yoichi Honda Masaaki Kuwahara 《Mycoscience》1996,37(4):417-425
Manganese peroxidase (MnP) and lignin peroxidase (LiP) were produced by growing a white-rot fungusBjerkandera adusta statically, on a wood meal/wheat bran culture in flasks. MnP and LiP reached their maximum activity after 6 and 19 days of
inoculation, respectively. Both MnP and LiP are thought to be important enzymes in lignin biodegradation byB. adusta. Ion exchange chromatography showed thatB. adusta produced a single LiP and a single MnP enzyme in wood meal/wheat bran culture. These enzymes were separated and characterized.
The molecular weight of MnP was 46,500 with a pl of 3.9. The molecular weight of LiP was estimated to be 47,000 with a pl of 3.5. Spectral analysis demonstrated that both enzymes are heme proteins. Production of these enzymes was also achieved
using a rotarysolid culture fermenter. MnP, LiP and veratryl alcohol oxidase were produced byB. adusta in the fermenter. 相似文献
143.
144.
A Ca2+-dependent protein kinase (CDPK) that has been partiallypurified and characterized previously [Yuasa and Muto (1992)Arch. Biochem. Biophys. 296: 175] was further purified to about20,000-fold from the soluble fraction of Dunaliella tertiolecta.The enzyme preparation contained 60- and 52-kDa polypeptidesboth of which phosphorylated casein as a substrate. Both polypeptidesshowed a Ca2+-dependent increase in mobility during SDS-PAGEand 45Ca2+-binding activity after SDS-PAGE and electroblottingonto a nitrocellulose membrane, suggesting that both the 60-and 52-kDa CDPKs directly bind Ca2+. The protein kinase inhibitors,K-252a and staurosporine, inhibited the CDPK competitively withrespect to ATP. An antibody raised against the 60-kDa CDPK crossreactedwith both the 60- and 52-kDa polypeptides. Both molecular specieswere autophosphorylated in the presence of Ca2+, and a highlyphosphorylated 80-kDa band appeared in addition to these phosphorylatedbands at 60 and 52 kDa in SDS-PAGE. However, the specific activityof CDPK was not changed by prior autophosphorylation when theautophosphorylated enzyme was assayed as a mixture of thesephosphorylated molecular species. Only the 60-kDa polypeptidewas immunodetected in subcellular fractions of Dunaliella cells.The 52-kDa polypeptide increased during storage of the enzyme.These results suggest that the 52-kDa polypeptide is a proteolyticartifact produced during purification. Immunoreactive bandsof 60-kDa were detected in extracts of several green algae butnot in extracts of higher plants or a brown alga.
1This research was partly supported by Grants-in-Aid from theMinistry of Education, Science and Culture, Japan (No. 06454013and 06304023) and Research Fellowship of the Japan Society forthe Promotion of Science for Young Sciencists.
2Research Fellow (PD) of the Japan Society for the Promotionof Science. 相似文献
145.
Kei-ichi Kuma Naruo Nikoh Naoyuki Iwabe Takashi Miyata 《Journal of molecular evolution》1995,41(2):238-246
The phylogenetic position of Dictyostelium inferred from 18S rRNA data contradicts that from protein data. Protein trees always show the close affinity of Dictyostelium with animals, fungi, and plants, whereas in 18S rRNA trees the branching of Dictyostelium is placed at a position before the massive radiation of protist groups including the divergence of the three kingdoms. To settle this controversial issue and to determine the correct position of Dictyostelium, we inferred the phylogenetic relationship among Dictyostelium and the three kingdoms Animalia, Fungi, and Plantae by a maximum-likelihood method using 19 different protein data sets. It was shown at the significance level of 1 SE that the branching of Dictyostelium antedates the divergence of Animalia and Fungi, and Plantae is an outgroup of the Animalia-Fungi-Dictyostelium clade.Correspondence to: T. Miyata 相似文献
146.
Changes in chorion proteins induced by the exudate released from the egg cortex at the time of fertilization in the teleost, Oryzias latipes 总被引:2,自引:2,他引:0
Takashi Iwamatsu Yasushi Shibata Toshihiro Kanie 《Development, growth & differentiation》1995,37(6):747-759
The chorion of unfertilized medaka Oryzias latipes eggs consists of two major proteins (77–73 and 49 kDa) and a minor 150 kDa protein. Upon fertilization, these major chorion proteins are polymerized to insoluble high molecular weight proteins via the temporary formation of several new proteins (132, 114, 62 and 61 kDa). Increasing chorion toughness is closely related to the formation of high molecular weight proteins and the increasing insolubility of the chorion proteins. The changes in chorion proteins and hardening could be induced in vitro in isolated chorions by an egg exudate, which includes cortical alveolar contents. The effects of temperature and pH on the egg exudate-induced changes in chorion proteins were examined in the present study. The major proteins could be digested by proteolytic enzymes. The 49 kDa protein was PAS-positive. Analysis with polyclonal antibodies against the major proteins demonstrated that the temporarily formed 62 and 61 kDa proteins were derived from the 77–73 kDa protein and that higher molecular weight proteins, newly formed in the process of chorion hardening, contained the same epitopes as did the 77–73 and 49 kDa proteins. The results suggest that the changes in chorion proteins of the medaka egg at the time of fertilization can be induced by an enzyme(s) released from the egg cortex into the perivitelline space. 相似文献
147.
Takashi Hirayarna Tatsuya Maeda Haruo Saito Kazuo Shinozaki 《Molecular & general genetics : MGG》1995,249(2):127-138
Yeast cells can respond and adapt to osmotic stress. In our attempt to clarify the molecular mechanisms of cellular responses to osmotic stress, we cloned seven cDNAs for hyperosmolarity-responsive (HOR) genes from Saccharomyces cerevisiae by a differential screening method. Structural analysis of the clones revealed that those designated HOR1, HORS, HOR4, HOR5 and HOR6 encoded glycerol-3-phosphate dehydrogenase (Gpd1p), glucokinase (Glklp), hexose transporter (Hxtlp), heat-shock protein 12 (Hsp12p) and Na+, K+, Li+-ATPase (Enalp), respectively. HOR2 and HOR7 corresponded to novel genes. Gpdlp is a key enzyme in the synthesis of glycerol, which is a major osmoprotectant in S. cerevisiae. Cloning of HOR1/GPD1 as a HOR gene indicates that the accumulation of glycerol in yeast cells under hyperosmotic stress is, at least in part, caused by an increase in the level of GPDH protein. We performed a series of Northern blot analyses using HOR cDNAs as probes and RNAs prepared from cells grown under various conditions and from various mutant cells. The results suggested that all the HOR genes are regulated by common signal transduction pathways. However, the fact that they exhibited certain distinct responses indicated that they might also be regulated by specific pathways in addition to the common pathways. Ca2+ seemed to be involved in the signaling systems. In addition, Hog1p, one of the MAP kinases in yeast, appeared to be involved in the regulation of expression of HOR genes, although its function seemed to be insufficient for the overall regulation of expression of these genes. 相似文献
148.
Tadashi II Masayuki Kubota Takashi Hirano Mamoru Ohashi Keiichi Yoshida Sakaru Suzuki 《Glycoconjugate journal》1995,12(3):282-289
The fast atom bombardment (FAB) collision induced dissociation (CID)-mass spectrometry/mass spectrometry (MS/MS) technique was successfully applied to characterize and identify the structures of the immunoreactive trisulfated and tetrasulfated tetrasaccharides that were obtained from the chondroitin sulfate in a shark fin using a treatment with chondroitinase ABC.Abbreviations FABMS
fast atom bombardment mass spectrometry
- CID
collision induced dissociation
- MS/MS
mass spectrometry/mass spectrometry
- UA2S-GalNAc6S
2-acetamido-2-deoxy-3-O-(2-O-sulfo--d-gluco-4-enepyranosyluronic acid)-6-O-sulfo-d-galactose
- UA-GalNAc4S
2-acetamido-2-deoxy-3-O-(-d-gluco-4-enepyranosyluronic acid)-4-O-sulfo-d-galactose
- UA-GalNAcDiS
2-acetamido-2-deoxy-3-O-(-d-gluco-4-enepyranosyluronic acid)-4,6-di-O-sulfo-d-galactose 相似文献
149.
Mutations at the flügellos (fl) locus in Bombyx mori give rise to wingless pupae and moths. To understand the developmental steps responsible for the fl wing defect, we compared the morphological changes and protein synthesis profiles between fl and wild-type (WT) wing discs during larval development. Morphologically, the four wing discs in the fl homozygote larva developed normally at least until the fourth instar, but they were slightly smaller than those of the WT.
After the last larval ecdysis, wing epithelial invagination and tracheal migration into the lacunar spaces evidently occurred
in the WT wing discs. However, there was no apparent morphological change in fl discs through the fifth instar. The fl wing discs cultured in medium containing 20-hydroxyecdysone (20E) did not grow and develop, although the WT wing discs extended
and differentiated under the same conditions. A comparison of protein synthesis in the wing discs revealed that several bands
were differentially expressed between the fl and WT. A 41-kDa band expressed abundantly from larval to pharate pupal stages in the WT wing discs was rarely observed in
fl discs. Furthermore, in vitro culture studies showed that the 41-kDa protein was induced by 20E and specifically synthesized
in WT wing discs after the wandering stage, but not in fl discs. The wing-specific protein synthesis and morphogenesis in fl wing discs may be blocked due to aberrant expression of the fl gene.
Received: 6 November 1996 / Accepted: 5 February 1997 相似文献
150.
Naruo Nikoh Naoyuki Iwabe Kei-ichi Kuma Mutsuhito Ohno Tsutomu Sugiyama Yoko Watanabe Kinya Yasui Zhang Shi-cui Katsuji Hori Yoshiro Shimura Takashi Miyata 《Journal of molecular evolution》1997,45(1):97-106
Previously we suggested that four proteins including aldolase and triose phosphate isomerase (TPI) evolved with approximately
constant rates over long periods covering the whole animal phyla. The constant rates of aldolase and TPI evolution were reexamined
based on three different models for estimating evolutionary distances. It was shown that the evolutionary rates remain essentially
unchanged in comparisons not only between different classes of vertebrates but also between vertebrates and arthropods and
even between animals and plants, irrespective of the models used. Thus these enzymes might be useful molecular clocks for
inferring divergence times of animal phyla. To know the divergence time of Parazoa and Eumetazoa and that of Cephalochordata
and Vertebrata, the aldolase cDNAs from Ephydatia fluviatilis, a freshwater sponge, and the TPI cDNAs from Ephydatia fluviatilis and Branchiostoma belcheri, an amphioxus, have been cloned and sequenced. Comparisons of the deduced amino acid sequences of aldolase and TPI from the
freshwater sponge with known sequences revealed that the Parazoa–Eumetazoa split occurred about 940 million years ago (Ma)
as determined by the average of two proteins and three models. Similarly, the aldolase and TPI clocks suggest that vertebrates
and amphioxus last shared a common ancestor around 700 Ma and they possibly diverged shortly after the divergence of deuterostomes
and protostomes. 相似文献