巻号一覧
110 巻, 1 号
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Gustavo A. Nores, Roger D. Dennis, Friedhelm Helling, Herbert Wiegandt1991 年 110 巻 1 号 p. 1-8
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーAs a consequence of detecting an IgM M-protein (naturally occurring diseased-state monoclonal antibody) immunoreactive to insect acidic glycolipids in a patient with demyelinating peripheral neuropathy, normal human sera were examined for the occurrence of heterophile antibodies directed against carbohydrate epitopes present on glyco-sphingolipids of Calliphora vicina (Insecta: Diptera). The insect glycolipids can be separated into neutral, zwitterionic, and acidic types, according to whether the oligosac-charide chains consist of neutral monosaccharides only, or carry an additional phospho-ethanolamine side chain and/or a β-glucuronic acid residue, respectively. Natural antibody activity to these three classes of insect glycosphingolipids was detected in all normal human sera examined. The antibody activities were separated by sequential chromatography on affinity columns of octyl-Sepharose 4B-bound neutral and zwitterionic glycolipids into three populations with differing epitope-type specificities. As expected for heterophile antibodies, they are mainly of the IgM class. Population I recognized epitopes present on the three types of insect glycolipids, i.e., the neutral oligosaccharide chain backbone, the main determinant of which contains a terminal N-acetylhexosamine. Immunoreactivity is separable into at least four subpopulations of differing carbohydrate epitope specificity. Population II recognized epitopes containing phosphoethanolamine in zwitterionic and some acidic insect glycolipids. There are two subpopulations, the majority of which require the free amino group of phosphoethanolamine for immunoreactivity. Population III antibodies showed immunoreactivity to terminal β-glucuronic acid-containing epitopes present only on acidic insect glycolipids.抄録全体を表示PDF形式でダウンロード (6000K) -
Hiroaki Kiyokawa, Hiroyuki Fukui, Hiroyuki Mizugruchi, Akiharu Kubo, N ...1991 年 110 巻 1 号 p. 9-11
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーAdenosine caused a 2.5-fold increase in the sodium-dependent uptake of 2-aminoisobutyric acid in rat hepatocytes in primary culture following incubation for 3 h. The range of stimulating concentrations of adenosine corresponded to that of cAMP formation. Adenosine increased the Vmax of the transport without altering the Km for 2-aminoisobutyric acid. These effects of adenosine were abolished by actinomycin D. N6-L-Phenylisopropyladenosine also caused twofold increase in the amino acid transport. These findings suggest that adenosine induces System A amino acid transport in a transcription-dependent manner, and that P1-purinergic receptors are mainly involved in this action of the nucleoside.抄録全体を表示PDF形式でダウンロード (1259K) -
Gerardo D. Fidelio, Toshio Ariga, Bruno Maggio1991 年 110 巻 1 号 p. 12-16
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーThe molecular area, collapse pressure, and surface potential of gangliosides obtained by different methods were systematically compared in monolayers at the air-water interface. Different values of these parameters are obtained depending on the purification procedure employed for the isolation of pure gangliosides. This is due to impurities (such as peptidaceous material) that remain in different amounts in the various preparations and that modify the ganglioside surface behavior. Routine purity checking by HPTLC analysis of gangliosides usually fails to reveal these impurities. On the other hand, even if the monolayer technique cannot identify the nature or amount of contaminants, it is extremely sensitive to reveal alterations of the surface molecular parameters caused by relatively small amounts of other components coextracted with the ganglioside or adventitiously introduced with the solvents or subphases employed. This is a serious problem for the obtention of correct and reproducible values of such important parameters as the molecular area of gangliosides, their electrostatic potential in oriented interfaces, and their interactions with other lipids and proteins. A procedure leading to consistent molecular parameters that remain reproducible after several repurification cycles is to perform an alkaline treatment on previously purified gangliosides species with NaOH, this is followed by dialysis against bidistilled water, rechromatography on DEAE-Sephadex A25, silicic acid or Iatrobeads, and Sephadex LH-20 columns; repurified gangliosides are stored in chloro-form-methanol-0.01M NaOH (60:30:4.5).抄録全体を表示PDF形式でダウンロード (698K) -
Setsu Kadowaki, Kenji Yamamoto, Masatoki Fujisaki, Tatsurokuro Tochiku ...1991 年 110 巻 1 号 p. 17-21
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーThe activity and substrate specificity of endo-β-N-acetylglucosaminidase [glycopeptide-D-mannosyl-N4-(N-acetyl-D-glucosaminy1)2-asparagine 1, 4-N-acetyl-β-glucosamino-hydrolase, EC 3.2.1.96] obtained from Mucor hiemalis (Endo-M) was compared with that of the enzyme obtained from Flavobacterium meningosepticum (Endo-F), which is the only enzyme available that acts on the complex oligosaccharides of asparagine-linked sugar chains in glycoproteins. They showed almost the same activities toward DNS-ovalbumin glycopeptide containing high-mannose and hybrid asparagine-linked oligosaccharides. However, Endo-M showed high activity towards DNS-asialotransferrin and DNS-transferrin glycopeptides, which contain complex biantennary oligosaccharides. Endo-M could weakly act even on DNS-asialofetuin glycopeptide containing complex triantennary oligosaccharides, while Endo-F could not. SDS-denatured asialotransferrin was deglycosylated by both enzymes in the presence of non-ionic detergent (NP-40) and EDTA, and the deglycosylated protein migrated to a lower molecular weight position than asialotrans-ferrin on SDS-PAGE. However, even in the absence of detergent, Endo-M deglycosylated native asialotransferrin and transferrin. Deglycosylation of asialotransferrin was confirmed by means of Con A-Sepharose 48 column chromatography and SDS-PAGE.抄録全体を表示PDF形式でダウンロード (4426K) -
Tadashi Shimamoto, Hiroshi Izawa, Hirohiko Daimon, Naotaka Ishiguro, M ...1991 年 110 巻 1 号 p. 22-28
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーA cryptic citrate transport gene (citA) from Salmonella typhimurium chromosome was cloned and its nucleotide sequence was determined. The cloned plasmid conferred citrate-utilizing ability on wild-type Escherichia coli, which cannot grow on citrate as the sole source of carbon. The resultant E. coli transformant was able to transport citrate. A 1, 302-base-pair open reading frame with a preceding ribosomal binding site was found in the cloned DNA fragment. The 434-amino-acid protein that could be translated from this open reading frame is highly hydrophobic (69% nonpolar amino acid residues), consistent with the fact that the transport protein is an intrinsic membrane protein. The molecular weight of this protein was calculated to be 47, 188. The gene sequence determined is highly homologous to those of Cit+ plasmid-mediated citrate transport gene, citA, from E. coli, the chromosomal citA gene from Citrobacter amalonaticus and the chromosomal cit+ gene from Klebsiella pneumoniae. The hydropathy profile of the deduced amino acid sequence suggests that this carrier has 12 hydrophobic segments, which may span the membrane lipid bilayer.抄録全体を表示PDF形式でダウンロード (853K) -
Hisashi Oku, Sumihiro Hase, Tokuji Ikenaka1991 年 110 巻 1 号 p. 29-34
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーAn α-mannosidase was purified from the magnum section of Japanese quail oviduct by
ammonium sulfate precipitation, DEAE-Sephacel chromatography, Sephacryl S-300 chromatography, mannan-Sepharose 4B chromatography, and hydroxyapatite chromatography. The purified α-mannosidase (referred to as neutral α-mannosidase) showed a single band on polyacrylamide gel with or without sodium dodecyl sulfate. Its molecular weight was found to be 330, 000 by gel chromatography. Neutral α-mannosidase hydrolyzed p-nitrophenyl α-D-mannopyranoside and the pyridylamino derivative of Man α1-6(Man α1-3)Man α1-6(Man α1-3)Man β1-4GlcNAc β1-4GlcNAc (Km value was 3mM). Mannosyl α1-2 linkages in the pyridylamino derivative of Manαl-2Manα1-6(Manαl-2Manαl-3)Manα1-6(Manαl-2Manα1-2Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAc were hardly hydrolyzed. Its optimum pH was found to be 7.0. The activity of the enzyme was activated by Co2+, and was potently inhibited by Cu2+, Hg2+, swainsonine, and 1-deoxymannojirimycin.抄録全体を表示PDF形式でダウンロード (1657K) -
Tomoko Iwata, Keiko Uchida-Mizuno, Takeshi Katafuchi, Teizo Ito, Hirom ...1991 年 110 巻 1 号 p. 35-39
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーType A atrial natriuretic peptide (ANP) receptor was demonstrated to be present as a tetramer in the bovine adrenal cortex. Type A ANP receptor is composed of two functional domains, namely extracellular ANP-binding and cytoplasmic guanylate cyclase domains, and generally considered to be present as a single polypeptide chain of about 140 kDa based on its primary structure deduced from the cDNA sequence and its SDS/PAGE profile under reducing conditions. Characterization of the type A receptor or receptor/cyclase under non-reducing conditions led to the discovery stated in the title. The type A ANP receptor was partially purified from bovine adrenal cortex membranes by Blue-Sepharose and GTP-agarose chromatography. SDS-PAGE analysis of the receptor preparation revealed that although under reducing conditions it migrated as a 140-kDa band, the mobility of the receptor was greatly retarded in the absence of reducing agents, suggesting that the type A ANP receptor is present as a disulfide-linked oligomer in its native state. Further analysis using SDS-polyacrylamide-agarose gels suitable for determining the sizes of high-molecular-weight proteins revealed that the oligomer has an Mr of 500, 000-550, 000. This result clearly indicates that the native form of the type A receptor is a tetramer composed of four 140-kDa disulfide-linked receptor/cyclase molecules.抄録全体を表示PDF形式でダウンロード (1882K) -
Yasushi Watanabe, Yoshiyuki Kijima, Masaaki Kadoma, Michihiko Tada, To ...1991 年 110 巻 1 号 p. 40-45
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーThe number of polypeptides constituting the oligomeric structure of canine phospholamban (a putative regulator of Ca2+ -ATPase of cardiac sarcoplasmic reticulum) stable even in the presence of sodium dodecyl sulfate was estimated through determination of the molecular weight of the oligomer. Owing to the small molecular size, the low UV-absorptivity and the limited availability, the molecular weight determination required very sophisticated application of the following technique, used as the only recourse: low-angle laser light scattering measurement combined with high-performance gel chromatography. The molecular weight of phospholamban oligomer was found to be 30, 400 and the number of subunits was concluded to be five after correction for the dependence of the apparent molecular weights on the protein concentration.抄録全体を表示PDF形式でダウンロード (780K) -
Shigeharu Harada, Kengo Kitadokoro, Takayoshi Kinoshita, Yasushi Kai, ...1991 年 110 巻 1 号 p. 46-49
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーA neutral protease, i.e., a zinc-containing metalloendoprotease from Streptomyces caespitosus, has been crystallized using acetone as a precipitating agent. The crystals diffract to better than 1.5 Å resolution when a rotating anode X-ray generator is used as an X-ray source. Protein phase angles were calculated by the multiple isomorphous replacement method using two heavy-atom derivatives (HgCl2 and CH3HgCl). A 6 Å resolution electron density map clearly showed molecular boundaries. Although its amino acid sequence is not known, the folding pattern of the polypeptide chain could be traced on a 2.5 Å resolution electron density map. A large cleft, which is located on the molecular surface, was proved to be the active site of the enzyme by structure analyses of inhibitor-complex crystals. The highest electron density peak, which corresponds to the cleft, was assigned to a catalytically essential zinc atom on difference Fourier synthesis between native and EDTA-soaked crystals.抄録全体を表示PDF形式でダウンロード (2577K) -
Hiroshi Yokota, Tomohiko Fukuda, Akira Yuasa1991 年 110 巻 1 号 p. 50-53
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーTwo isoforms of UDP-glucuronyltransferase purified from rat liver (named GT-1) and kidney (named GT-2) have various properties in common but differ in their NH2-terminal sequences. In this study, the two forms were further found to have common immuno-chemical properties, i.e., they could not be distinguished by Ouchterlony double diffusion and immunoblotting analyses. These isoforms also had the same inducibility as shown by immunoblotting analysis: GT-2 protein in rat was increased by treatment with β-naphtho-fiavone and 3-methylcholanthrene, whereas GT-1 was inducible by 3-methylcholanthrene. However, the effects of phospholipids on these enzymes were extremely different. 1-Naphthol glucuronizing activity of GT-1 was increased 7.5-8-fold by lysophosphatidyl-choline, but the activity of GT-2 was increased only 3-3.6-fold. The transferase activity of GT-1 toward 4-methylumbelliferone was increased 2-2.5-fold by dilauroylphosphatidyl-choline, but that of GT-2 was reduced, while its 4-nitrophenol glucuronidation activity was increased 1.5-fold by the phospholipid. These results indicate that the two similar UDP-glucuronyltransferases from rat liver and kidney interact differently with phos-pholipids and that the activation level of UDP-glucuronyltransferase activity with phos-pholipids depends on the aglycone substrates.抄録全体を表示PDF形式でダウンロード (1854K) -
Masaki Hayashida, Tetsuo Maita, Genji Matsuda1991 年 110 巻 1 号 p. 54-59
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーSubfragment-1 was prepared from adult chicken pectoralis myosin by limited digestion with α-chymotrypsin, and an amino-terminal 23 kDa fragment of the heavy chain was obtained by digesting the subfragment-1 with trypsin. The 205-residue sequence of the fragment was determined by sequencing its cyanogen bromide, tryptic, and chymotryptic peptides. The amino-terminal α-amino group of the fragment was acetylated, and two methylated lysines; ε-N-monomethyllysine and ε-N -trimethylly sine were recognized at the 35 th and 130th positions, respectively, as in rabbit skeletal myosin. Comparing the 205-residue sequence of the skeletal myosin with those of cardiac, and gizzard myosins from chicken, considerable differences are recognized, especially in the amino-terminal region, but strong homologies are observed around the reactive lysine residue, around the ε-N-trimethyllysine residue, and around the consensus sequence of GXXGXGKT for nucleotide-binding proteins. On the other hand, only 12 amino acid substitutions are recognized between adult and embryonic skeletal myosins, allowing for the post-translational methylation.抄録全体を表示PDF形式でダウンロード (690K) -
Yoko Komine, Tetsuo Maita, Genji Matsuda1991 年 110 巻 1 号 p. 60-67
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーThe complete amino acid sequence of the 50 kDa fragment of subfragment-1 from adult chicken pectoralis muscle myosin was determined. It contained 431 residues including an ε-N-trimethyllysine at position 346. The 431-residue sequence corresponds to the sequence of residues 206 to 639 of chicken embryonic breast muscle myosin heavy chain which was predicted from the nucleotide sequence of the cDNA by Molina et al. [Molina, M.I., Kropp, K.E., Gulick, J., & Robbins, J. (1987) J. Biol. Chem. 262, 6478-6488]. Comparing the two sequences, 23 amino acid substitutions and three deletions/insertions are recognized.抄録全体を表示PDF形式でダウンロード (779K) -
Tetsuo Maita, Takayuki Miyanishi, Kazuhisa Matsuzono, Yoshito Tanioka, ...1991 年 110 巻 1 号 p. 68-74
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーThe amino acid sequence of the 197-residue 22 kDa fragment from chicken pectoralis muscle was determined to be as follows: K-K-G-S-S-F-Q-T-V-S-A-L-F-R-E-N-L-N-K-L-M-A-N-L-R-S-T-H-P-11-F-V-R-C-I-I-P-N-E-T-K-T-P-G-A-M-E-H-E-L-V-L-H-Q-L-R-C-N-G-V-L-E-G-I-R-I-C-R-K-G-F-P-S-R-V-L-Y-A-D-F-K-Q-R-Y-R-V-L-N-A-S-A-I-P-E-G-Q-F-M-D-S-K-K-A-S-E-K-L-L-G-S-I-D-V-D-h-T-Q-Y-R-F-G-H-T-K-V-F-F-K-A-G-L-L-G-L-L-E-E-M-R-D-D-K-L-A-E-I-I-T-R-T-Q-A-R-C-R-G-F-L-M-R-V-E-Y-R-R-M-V-E-R-R-E-S-I-F-C-I-Q-Y-N-V-R-S-F-M-N-V-K-11-W-P-W-M-K-L-F-F-K, where h stands for 3-N-methylhistidine. The amino acid sequences of the 22 kDa fragment and its equivalent fragment from chicken ventricle and gizzard muscle myosins were also determined by our group. Predicted secondary structures of these 22 kDa fragment regions and of the reported chicken embryo myosin revealed some possible structural differences. There is a highly conserved 15-residue region in the vicinity of reactive cysteine residues, a region which has been suggested to bind actin and ATP analogue. This possible functional region may have a unique secondary structure containing a turn or coil structure.抄録全体を表示PDF形式でダウンロード (2226K) -
Tetsuo Maita, Eiko Yajima, Shuichi Nagata, Takayuki Miyanishi, Susumu ...1991 年 110 巻 1 号 p. 75-87
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーIn the preceding paper [Maita, T., Miyanishi, T., Matsuzono, K., Tanioka, Y., & Matsuda, G. (1991) J. Biochem. 110, 68-74], we reported the amino-terminal 837-residue sequence of the heavy chain of adult chicken pectoralis muscle myosin. This paper describes the carboxyl terminal 1, 097-residue sequence and the linkage of the two sequences. Rod obtained by digesting myosin filaments with α-chymotrypsin was redigested with the protease at high KC1 concentration, and two fragments, subfragment-2 and light meromyosin, were isolated and sequenced by conventional methods. The linkage of the two fragments was deduced from the sequence of an overlapping peptide obtained by cleaving the rod with cyanogen bromide. The rod contained 1, 039 amino acid residues, but lacked the carboxyl-terminal 58 residues of the heavy chain. A carboxyl-terminal 63-residue peptide obtained by cleaving the whole heavy chain with cyanogen bromide was sequenced. Thus, the carboxyl terminal 1, 097-residue sequence of the heavy chain was completed. The linkage of subfragment-1 and the rod was deduced from the sequence of an overlapping peptide between the two which was obtained by cleaving heavy meromyosin with cyanogen bromide. Comparing the sequence of the adult myosin thus determined with that of chicken embryonic myosin reported by Molina et al. [Molina, M.I., Kropp, K.E., Gulick, J., & Robbins, J. (1987) J. Biol. Chem. 262, 6478-6488], we found that the sequence homology is 94%.抄録全体を表示PDF形式でダウンロード (2122K) -
Kiyoshi Ikeda, Seiji Inoue, Chika Amasaki, Keizo Teshima, Hiroh Ikezaw ...1991 年 110 巻 1 号 p. 88-95
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーThe phosphatidylcholine-hydrolyzing phospholipase C, so-called “phospholipase C” (PLC), was isolated from the culture of Bacillus cereus strain IAM 1208. The amino-acid composition and partial N-terminal sequence of the purified enzyme were in good agreement with those expected from the nucleotide sequence for a PLC of strain ATCC 10987 [Johansen et al. (1988) Gene 65, 293-304]. The chain-length dependence of kinetic parameters for the PLC-catalyzed hydrolysis of monodispersed short-chain phosphatidylcholines (diCNPC, N=3-6) was studied by a pH-stat assay method at 25°C, pH 8.0, and ionic strength 0.2 in the presence of saturating amounts of Zn2+ (0.1mM). The result was compared with those for snake venom phospholipases A2 [Teshima et al. (1989) J. Biochem. 106, 518-527]. It was found that the interaction of the PLC with the head group of the substrate molecule is very important for the binding. The pH dependences of kinetic parameters for the hydrolysis of monodispersed diC5PC and mixed micelles of diC16PC with Triton X-100 were also studied under the same conditions. An ionizable group, whose pK value is perturbed from 7.77 to 8.30 by substrate binding, was found to be essential to the catalysis. This group was tentatively assigned to His 14 on the basis of the results on X-ray crystallographic and chemical modification studies [Hough et al. (1989) Nature 338, 357-360 and Little (1977) Biochem. J. 167, 399-404]. Another ionizable group, with a pK value of 7.60, which participates slightly in the catalysis, was thought to be the α-amino group located in close proximity to His 14. A third ionizable group, with a pK value of 5.33, was also found to be essential to the catalysis. This group was tentatively assigned to Glu 146, to which a Zn2+ ion loosely coordinates in the active site抄録全体を表示PDF形式でダウンロード (889K) -
Kei-ichi Uemura, Eiko Sugiyama, Tamotsu Taketomi1991 年 110 巻 1 号 p. 96-102
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリー1-Phenyl-2-decanolyamino-3-morpholino-l-propanol (PDMP), an effective inhibitor of UDP-glucose:ceramide glucosyltransferase, caused inhibition of cell growth in murine neuroblastoma cell lines. Metabolic labeling of glycosphingolipids with [14C]galactose in NS-20Y, Neuro2a, and N1E-115 cells showed reduced incorporation of radioactivity into gangliosides and neutral glycosphingolipids when threo-PDMP was present in the medium. Treatment of NS-20Y cells with threo-PDMP resulted in a time-dependent decrease in mass levels of gangliosides and neutral glycosphingolipids. After 24 h in the presence of 50 μM threo-PDMP, neutral glycosphingolipid mass was reduced to 32%, where glucosylceramide was the most affected (90% decrease). The ganglioside mass was reduced to 57% of the original content. Neurite outgrowth from neuroblastoma cells in serum-free medium was significantly inhibited by threo-PDMP in a dose-dependent manner. Threo-PDMP also caused retraction of neurites which had been induced to extend in serum-free medium. Pretreatment of cells with GM1 partially restored the ability of NS-20Y cells for neurite outgrowth in the medium containing threo-PDMP. These results suggest a possible role for glycosphingolipids in neurite outgrowth of murine neuroblastoma cells.抄録全体を表示PDF形式でダウンロード (2689K) -
Ken Okabayashi, Yukimitsu Nakagawa, Naofumi Hayasuke, Hideyuki Ohi, Ma ...1991 年 110 巻 1 号 p. 103-110
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーWe have fused a cDNA gene encoding mature human serum albumin (HSA) to several secretory leader-encoding sequences. The hybrid genes were cloned into an episomal vector under the control of several yeast promoters and then introduced into yeast cells. The GAL1 promoter in combination with either the native HSA pre-sequence or a modified HSA pre-sequence gave the highest production of immunoreactive HSA, 90mg/liter being reached in a shake flask culture. The invertase pre-sequence, the mating factor α1 prepro-sequence, and the modified HSA pre-sequence directed accurate processing. In contrast, the chicken lysozyme pre-sequence and the native HSA pre-sequence directed incorrect processing. Episomal vectors were unstable within the host cells under non-selective culture conditions. To improve the plasmid stability, the hybrid genes were incorporated into an integrative vector. Transformants carrying multicopies of the plasmid integrated at the LEU2 locus stably secreted HSA. The highest yield of 65mg/liter in a shake flask culture was obtained with the combination of the yeast glyceraldehyde-3-phosphate dehydrogenase promoter and the modified HSA pre-sequence. By constructing transformed strains containing multicopies of plasmids integrated at both the chromosome LEU2 and HIS4 loci, we have obtained a stable strain that continuously secretes as much as 85mg HSA per liter of culture medium.抄録全体を表示PDF形式でダウンロード (2159K) -
Kazuko Hori, Yoshihiro Yamamoto, Kazuhiko Tokita, Fumihiko Saito, Tosh ...1991 年 110 巻 1 号 p. 111-119
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーA fragment encoding proline-activating domain (grs 2-pro) of gramicidin S synthetase 2 (GS 2) was found in an 8.1-kilobase pairs (kb) DNA fragment of Bacillus brevis Nagano, which contained the full length of GS 1 gene (grs 1). The clones designated GS719 and GS708, which expressed gramicidin S synthetase 1, were elucidated to express immunoreactive proteins to GS 2 antibodies with approximate molecular weights of 115, 000, 105, 000 (GS719), and 110, 000 (GS708). The partial purification of the gene products of these clones was carried out using DEAE-Sepharose CL-6B column chromatography. The immunoreactive proteins to GS 2 antibodies were separated from gramicidin S synthetase 1 protein and had specific proline-dependent ATP-32PP1 exchange activity. The nucleotide sequence for the proline-activating domain in the 8.1-kb insert was determined. This fragment was 2, 879 base pairs long, and encoded 959 amino acids. The calculated molecular weight of 111, 671 was consistent with the apparent molecular weight of 115, 000 found in SDS-PAGE of the immunoreactive products to GS 2 antibodies. The open reading frame for this protein followed grs 1 gene, though two were separated by a 73-base pair noncoding sequence, and remained open to the end. A comparison of the putative amino acid sequence of grs 2-progene product showed that the amino-terminal region was highly homologous to the available sequence of tyrocidine synthetase 2 (71% similarity) and the carboxy-terminal region of the gene product had significant homology to the amino-terminal half of GS1 (66% similarity), as well as to other antibiotic peptide synthetases, tyrocidine synthetase 1 and δ-(L-α-aminoadipy1)-L-cysteinyl-D-valine synthetase (ACVS), an enzyme involved in penicillin biosynthesis.抄録全体を表示PDF形式でダウンロード (2124K) -
Jan Holgersson, Per-Åke Jovall, Michael E. Breimer1991 年 110 巻 1 号 p. 120-131
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーNon-acid glycosphingolipid expression was studied in the large intestines from four individuals with the A1Le(a-b+), BLe(a-b+), and OLe(a-b+) blood group phenotypes. In the A1Le(a-b+) case, specimens were taken from the ascending and sigmoid parts of the large intestine in order to compare the expression of glycolipids in the proximal and distal regions of the intestine. In one blood group Ole(a-b+) individual, epithelial cells were isolated from the residual stroma to compare the glycolipid compositions in these two tissue compartments. GlcCer, GalCer, LacCer, Gb3Cer, and Gb4Cer were the major compounds in all three individuals, as shown by mass spectrometry, proton NMR spectroscopy, and degradation studies. The Lea-5 glycolipid was the major complex blood group glycolipid in all individuals, except in the proximal ascending part of the large intestine of the A1Le(a-b+) case, in which the Leb-6 glycolipid was predominant. There were trace amounts of blood group ABH glycolipids, in agreement with the ABO blood group phenotypes of the donors, Lewis antigens with more than six sugar residues in the carbohydrate chain, and blood group X and Y glycolipid antigens. The epithelial cells were dominated by monoglycosylceramides and the Lea-5 glycolipid, while only trace amounts of di-, tri-, and tetraglycosylceramide structures were present. No reactivity was seen in the epithelial cell fraction with Galα1-4Gal specific Escherichia coli, anti-Pk, or anti-P antibodies, indicating the absence of the glycolipid-borne Galα1-4Gal sequence in human large intestinal epithelial cells.抄録全体を表示PDF形式でダウンロード (5056K) -
Atsushi Kon, Keiichi Takagaki, Hitoshi Kawasaki, Toshiya Nakamura, Mas ...1991 年 110 巻 1 号 p. 132-135
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーA pyridylamination method was applied to glycosaminoglycans and the characteristics of the resulting pyridylamino glycosaminoglycans were examined. First, glycosaminoglycan chains, which uniformly possess a xylose residue at their reducing termini, were liberated from proteoglycan by successive digestion with protease and endo-β-xylosidase. Then the glycosaminoglycan chains were coupled with 2-aminopyridine by reductive amination with sodium cyanoborohydride for 15 h according to the method of Hase, S. et al. [J. Biochem. 95, 197-203 (1984)]. The pyridylamination reaction caused neither depolymerization, de-N-acetylation, nor de-N-or de-O-sulfation. The pyridylamino glycosaminoglycan chains had an intact linkage region (GlcA-Gal-Gal-Xyl) between the carbohydrate chain and the peptide core of the proteoglycan. These pyridylamino glycosaminoglycans should be useful as substrates for endo-type glycosidases that act on glycosaminoglycan chains and as markers for studies of glycosaminoglycan metabolism.抄録全体を表示PDF形式でダウンロード (1424K) -
Seiko Shigeta, Kazuhisa Ono, Satoru Oka1991 年 110 巻 1 号 p. 136-140
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーA β-galactosidase was extracted from the internal organs of a sea squirt, Styela plicata, and purified 959-fold, with an 18% yield, by successive gel chromatography, anion-ex-change chromatography, chromatofocusing, and affinity chromatography on a Con A-Sepharose column. The purified enzyme was fairly homogeneous, as judged on disc PAGE, SDS-PAGE, and gel chromatography on a Sephadex G-200 column. The molecular weight of the enzyme was estimated to be 77, 000 and 75, 000 by gel chromatography and SDS-PAGE, respectively, and its isoelectric point was determined to be 4.9 by the isoelectric focusing method. The enzyme was substantially stable in the pH range of 3.5 to 7.5, the optimum pH being 4.0. The enzyme was significantly inhibited by 9mM HgCl2 and 9mM DFP, while the inhibition by 0.9% PCMB was only 60% at 0°C for 30min. The purified β-galactosidase apparently liberated galactose from a sea squirt antigen (H-antigen), two allergenically active glycopeptides (Gp-1 and Gp-2) derived from another sea squirt antigen (Gi-rep), asialo-ovomucoid glycopeptide, asialo-fetuin glycopeptide, GA1, CDH, and an ABEE-derivative (Galβ1→3ThrNAc-ABEE) of Ga1β1→3Ga1NAc-ol isolated from bovine submaxillary gland mucin.抄録全体を表示PDF形式でダウンロード (1402K) -
Masahide Yamamoto, Yoshiaki Takahashi, Kouichi Inano, Tsuneyoshi Horig ...1991 年 110 巻 1 号 p. 141-145
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーThe modes of binding of heat shock protein 90 with phenyl-Sepharose, myristoylatec AE-cellulose, and monomyristoylated lysozyme were studied to characterize a hydrophobic region(s) on the surface of the heat shock protein 90 molecule and the following results were obtained. (1) The binding of heat shock protein 90 with phenyl-Sepharose was inhibited by the addition of 30% ethylene glycol. This indicates that the binding involves a hydrophobic interaction. (2) The binding was strengthened by the addition of 10mM Mg2+, Ca2+, Sr2+ and Ba2+ ions, but not by K+ or Na+ ions. (3) The binding of hsp 90 with phenyl-Sepharose decreased initially and then increased as the temperature was increased from 0 to 50°C, with a minimum at around 35°C. (4) Lowering the pH stimulated the binding of hsp 90 with phenyl-Sepharose. (5) Heat shock protein 90 bound to myristoylated AE-cellulose, which has aliphatic hydrophobic residues, but not to acetylated AE-cellulose. (6) Heat shock protein 90 bound to monomyristoylated lysozyme, but not to control unmodified lysozyme. Based on these results, the possible function of the hydrophobic region(s) of heat shock protein 90 in the interaction with hydrophobic proteins is discussed.抄録全体を表示PDF形式でダウンロード (573K) -
Tadamasa Murakami, Hisamitsu Ohmori, Sadao Gotoh, Tohru Tsuda, Ryoichi ...1991 年 110 巻 1 号 p. 146-150
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーCultured human neuroblastoma (GOTO) cells were induced to differentiate by dibutyryl cyclic AMP (Bt2cAMP) and/or retinoic acid (RA). A combination of Bt2cAMP (1mM) and RA (1 μM) yielded the most significant networks of neurites after 3 to 4 days, this being associated with the reduction of N-mye mRNA levels. Next, we examined several cellular genes that were possibly linked with changes in N-myc gene expression under these conditions. Among the genes examined, both nucleolin and a major heat-shock protein (hsp70) mRNAs showed changes concomitant with those in N-myc mRNA levels when induced by Bt2cAMP and RA. Dibutyryl cAMP alone induced several short cellular processes and caused a marked decrease in N-myc mRNA within 2 days. RA alone induced a few long and straight neurites along the longitudinal axis of individual cells and a significant decrease in growth rate but showed neither network formation nor a decrease in N-myc gene expression. These results indicate differential effects of Bt2cAMP and RA on the regulatory mechanisms of both cell proliferation and differentiation and also indicate a possible association of expression of N-myc gene with those of hsp70 and nucleolin genes.抄録全体を表示PDF形式でダウンロード (2994K) -
Akihiro Iwamatsu, Hideyuki Aoyama, Gábor Dibó, Susumu Ts ...1991 年 110 巻 1 号 p. 151-158
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーThe amino acid sequence of nuclease S1, a nuclease which cleaves both single-stranded DNA and RNA, from Aspergillus oryzae was determined. Reduced and S-carboxymethylated or S-aminoethylated nuclease Si was digested with Achromobacter protease I, Staphylococcus aureus V8 protease, or endoproteinase Asp-N. Peptides thus obtained were purified by reverse-phase high-performance liquid chromatography and sequenced, and the complete primary structure was established. Nuclease Si consists of a single peptide chain of 267 amino acid residues bearing N-glycosylated Asns 92 and 228. Five half-cystine residues are present at positions 25, 72, 80, 85, and 216, and the latter four residues are implicated in the formation of disulfide bonds by analogy with those in nuclease P1. Two short stretches of sequences involving His 60 and His 125 are shown to be identical with those involving active site His 119 in bovine ribonuclease A and active-site His 134 in porcine deoxyribonuclease I, respectively.抄録全体を表示PDF形式でダウンロード (870K) -
Hideki Nakagoshi, Yoshio Ueno, Fumio Imamoto1991 年 110 巻 1 号 p. 159-162
発行日: 1991年
公開日: 2008/11/18
ジャーナル フリーThree prokaryotic termination signals, the Escherichia coli trp attenuator, λ4S RNA terminator, and λtR1 terminator, were examined as to their effects on transcription in vivo in HeLa cells. The trp attenuator inhibited the expression of downstream genes in an orientation-dependent manner, but both the λ4S RNA and λtR1 terminators did not. Furthermore, a point mutation of the attenuator that disrupted the secondary structure of mRNA abolished this inhibitory effect. These results suggest that an attenuator-like secondary structure is effective in inhibiting transcriptional elongation by RNA polymer-ase II.抄録全体を表示PDF形式でダウンロード (2932K)
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