Misc - 高橋　素子

家族性筋萎縮性側索硬化症に関連したCu,Zn-SOD変異体のフルクトースによる糖化の亢進

高宮 里奈, 高橋 素子, 宮澤 伸子, 藤原 範子, 宮本 泰豪, 谷口 直之

生化学 ( (公社)日本生化学会 )  73 ( 8 ) 1070 - 1070  2001年08月

N-アセチルグルコサミン転移酵素III(GnT-III)発現細胞における過酸化水素によるアポトーシスの抑制機構

渋川 幸直, 高橋 素子, 本家 孝一, 谷口 直之

生化学 ( (公社)日本生化学会 )  73 ( 8 ) 977 - 977  2001年08月

Overexpression of N-acetylglucosaminyltransferase III enhances the epidermal growth factor-induced phosphorylation of ERK in HeLaS3 cells by up-regulation of the internalization rate of the receptors

Y Sato, M Takahashi, Y Shibukawa, SK Jain, R Hamaoka, J Miyagawa, Y Yaginuma, K Honke, M Ishikawa, N Taniguchi

JOURNAL OF BIOLOGICAL CHEMISTRY ( AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC )  276 ( 15 ) 11956 - 11962  2001年04月

　概要を見る

N-Acetylglucosaminyltransferase III (GnT-III) is a key enzyme that inhibits the extension of N-glycans by introducing a bisecting N-acetylglucosamine residue. In this study we investigated the effect of GnT-III on epidermal growth factor (EGF) signaling in HeLaS3 cells. Although the binding of EGF to the epidermal growth factor receptor (EGFR) was decreased in GnT-III transfectants to a level of about 60% of control cells, the EGF-induced activation of extracellular signal-regulated kinase (ERR) in GnT-III transfectants was enhanced to similar to1.4-fold that of the control cells. A binding analysis revealed that only low affinity binding of EGF was decreased in the GnT-III transfectants, whereas high affinity binding, which is considered to be responsible for the downstream signaling, was not altered, EGF-induced autophosphorylation and dimerization of the EGFR in the GnT-III transfectants were the same levels as found in the controls. The internalization rate of EGFR was, however, enhanced in the GnT-III transfectants as judged by the uptake of I-125-EGF and Oregon Green-labeled EGF. Then the EGFR internalization was delayed by dansylcadaverine, the up-regulation of ERK phosphorylation in GnT-III transfectants was completely suppressed to the same level as control cells. These results suggest that GnT-III overexpression in HeLaS3 cells resulted in an enhancement of EGF-induced ERR phosphorylation at least in part by the upregulation of the endocytosis of EGFR.

DOI PubMed CiNii

N-アセチルグルコサミン転移酵素(GnT-III)導入HeLaS3細胞におけるEGFシグナリング

佐藤 祐一, 高橋 素子, 柳沼 裕二, 石川 睦男, 谷口 直之

日本癌学会総会記事 ( 日本癌学会 )  59回   486 - 487  2000年09月

N-アセチルグルコサミン転移酵素III(GnT III)発現細胞のアポトーシス抑制機構

渋川 幸直, 高橋 素子, 佐藤 祐一, 本家 孝一, 谷口 直之

生化学 ( (公社)日本生化学会 )  72 ( 8 ) 929 - 929  2000年08月

糖転移酵素GnT-III過剰発現細胞におけるEGFシグナリング

高橋 素子, 佐藤 祐一, 渋川 幸直, 本家 孝一, 谷口 直之

生化学 ( (公社)日本生化学会 )  72 ( 8 ) 983 - 983  2000年08月

Osmotic stressによるヘパリン結合成EGF様増殖因子(HB-EGF)の発現誘導と作用機序

高 永鎬, 車 文一, 鈴木 敬一郎, 朴 用軾, 高橋 素子, 東山 繁樹, 谷口 直之

生化学 ( (公社)日本生化学会 )  72 ( 8 ) 907 - 907  2000年08月

家族性筋萎縮性側索硬化症における変異Cu,Zn-SODの糖化反応の亢進

高宮 里奈, 宮本 泰豪, 高橋 素子, Theingi Myint, 宮澤 伸子, 谷口 直之

生化学 ( (公社)日本生化学会 )  72 ( 8 ) 1064 - 1064  2000年08月

Cloning of amadoriase I isoenzyme from Aspergillus sp.: Evidence of FAD covalently linked to Cys342

XL Wu, M Takahashi, SG Chen, VM Monnier

BIOCHEMISTRY ( AMER CHEMICAL SOC )  39 ( 6 ) 1515 - 1521  2000年02月

　概要を見る

Amadoriases are a novel class of FAD enzymes which catalyze the oxidative deglycation of glycated amino acids to yield corresponding amino acids, glucosone, and H2O2. We previously reported the purification and characterization of two amadoriase isoenzymes from Aspergillus fumigatus and the molecular cloning of amadoriase II. To identify the primary structure of amadoriase I, we prepared a cDNA library from Aspergillus fumigatus and isolated a clone using a probe amplified by polymerase chain reaction with primers designed according to the partial amino acid sequences from peptide mapping. The primary structure of the enzyme deduced from the nucleotide sequence comprises 445 amino acid residues. The enzyme contains 1 mol of FAD as a cofactor, which is covalently linked to Cys342, as determined by mutagenesis analysis, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and electrospray ionization-collisional-activated dissociation tandem mass spectrometry. Sequence alignment studies show that amadoriase I has 22% homology with monomeric sarcosine oxidase in which FAD is also linked to a homologous Cys residue. Amadoriases are of potential importance as tools for uncoupling hyperglycemia and glycation reactions that are thought to play a role in diabetic complications.

DOI PubMed CiNii

医学研究 In vitro kinaseアッセイ

高橋 素子

THE LUNG-perspectives ( (株)メディカルレビュー社 )  7 ( 4 ) 429 - 431  1999年10月

EGFレセプターに対する糖鎖付加の機能,構造への影響

津田 岳夫, 池田 義孝, 山口 幸洋, 盛 音, 井原 秀之, 高橋 素子, 谷口 直之

生化学 ( (公社)日本生化学会 )  71 ( 8 ) 1054 - 1054  1999年08月

糖転移酵素GnT-III導入CHO細胞におけるEGF受容体の構造変化

高橋 素子, 渋川 幸直, 井原 祥一, 浜岡 理恵子, 池田 義孝, 谷口 直之

生化学 ( (公社)日本生化学会 )  71 ( 8 ) 853 - 853  1999年08月

糖転移酵素GnT-III導入HeLaS3細胞におけるEGF刺激による細胞内情報伝達

佐藤 祐一, 高橋 素子, Jain Suresh K, 浜岡 理恵子, 池田 義孝, 石川 睦男, 谷口 直之

生化学 ( (公社)日本生化学会 )  71 ( 8 ) 854 - 854  1999年08月

Overexpression of the aldose reductase gene induces apoptosis in pancreatic beta-cells by causing a redox imbalance

R Hamaoka, J Fujii, J Miyagawa, M Takahashi, M Kishimoto, M Moriwaki, K Yamamoto, Y Kajimoto, Y Yamasaki, T Hanafusa, Y Matsuzawa, N Taniguchi

JOURNAL OF BIOCHEMISTRY ( JAPANESE BIOCHEMICAL SOC )  126 ( 1 ) 41 - 47  1999年07月

　概要を見る

To determine the role of the polyol metabolizing pathway under hyperglycemic conditions, the effects of aldose reductase (AR) on the cellular functions of pancreatic beta-cells were examined. Stable transfectants of rat AR cDNA were obtained with a pancreatic beta-cell line, HIT, in which a negligible amount of AR was originally expressed. Overproduction of AR triggered DNA fragmentation, as judged with the TUNEL method and agarose gel electrophoresis, Morphological analysis by electron microscopy also clearly showed apoptosis of the AR-overexpressing HIT cells, Induction by interleukin-1 beta of gene expression such as those of an inducible form of nitric oxide synthase (NOS-II) and Mn-superoxide dismutase (Mn-SOD), was much lower in the transfectants than in the control cells, while the expression of constitutively expressed genes such as those for Cu,Zn-superoxide dismutase and insulin was not changed. The susceptibility to interleukin-1 beta stimulation of the expression of the NOS II and Mn-SOD genes was due to suppressed NF-kappa B activity, which is essential for the expression of these genes. In addition, the intracellular NADPH/NADP(+) ratio was considerably lower in the AR-transfected cells than in control cells. Thus, the overexpression of AR in pancreatic beta-cells induced apoptosis that may be caused by a redox imbalance.

Mn-superoxide dismutase(Mn-SOD)のミトコンドリア移行の変異体を用いた解析

藤井 順逸, 池田 義隆, 渡辺 俊哉, 川崎 佳巳, 高橋 素子, 鈴木 敬一郎, 谷口 直之

生化学 ( (公社)日本生化学会 )  70 ( 8 ) 988 - 988  1998年08月

A defect in the mitochondrial import of mutant Mn-superoxide dismutase produced in Sf21 cells

J Fujii, Y Ikeda, T Watanabe, Y Kawasaki, K Suzuki, C Fujii, M Takahashi, N Taniguchi

JOURNAL OF BIOCHEMISTRY ( OXFORD UNIV PRESS )  124 ( 2 ) 340 - 346  1998年08月

　概要を見る

Wild-type and several mutant human manganese superoxide dismutases (Mn-SODs) were produced in a baculovirus/insect cell system and characterized. The enzymatic activity of a homogenate of Sf21 cells, infected with baculovirus carrying wild-type Mn-SOD and grown in the conventional medium, was indistinguishable from that of control cells, but was augmented by supplementation with Mn2+. The protein produced was largely imported into the mitochondria, as judged from the enrichment in the mitochondrial fraction, the mobility of the protein on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the results of N-terminal processing, which was confirmed by sequencing of the purified enzyme. However, a significant amount of precursor was also detected by an antibody raised against the human Mn-SOD signal peptide, While both Mn2+ and Fe3+ stimulated Mn-SOD accumulation within mitochondria, the active form was produced in the presence of submillimolar Mn2+ only. Amino acid substitutions at a signal peptide-cleavage site, His-Ser-Leu(+) to Pro-Met-Val(+), in the mature Mn-SOD prevented the processing of the precursor protein, and thus resulted in the accumulation of the precursor protein within mitochondria, as judged on immunostaining with an anti-Mn-SOD antibody. Mutant Mn-SODs with a truncated signal peptide or carboxyl region (8, 13, and 42 amino acid residues in the mature form) were barely solubilized, even with a nonionic detergent, and exhibited no activity, suggesting inappropriate folding of these mutant SODs, They were also susceptible to proteolytic degradation, while the wild-type and precursor forms were resistant, Thus, the baculovirus/insect cell expression system appears to be adequate for the analysis of mitochondrial import using intact cells as well as for the large scale production of active Mn-SOD.

Selective induction of heparin-binding epidermal growth factor-like growth factor by methylglyoxal and 3-deoxyglucosone in rat aortic smooth muscle cells - The involvement of reactive oxygen species formation and a possible implication for atherogenesis in diabetes

WY Che, M Asahi, M Takahashi, H Kaneto, A Okado, S Higashiyama, N Taniguchi

JOURNAL OF BIOLOGICAL CHEMISTRY ( AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC )  272 ( 29 ) 18453 - 18459  1997年07月

　概要を見る

Methylglyoxal (MG) and 3-deoxyglucosone (3-DG), reactive dicarbonyl metabolites in the glyoxalase system and glycation reaction, respectively, selectively induced heparin-binding epidermal growth factor (HB-EGF)-like growth factor mRNA in a dose- and time-dependent manner in rat aortic smooth muscle cells (RASMC). A nuclear run-on assay revealed that the dicarbonyl may regulate expression of HB-EGF at the transcription level. The dicarbonyl also increased the secretion of HB-EGF from RASMC. However, platelet-derived growth factor, another known growth factor of smooth muscle cells (SMC), was not induced by both dicarbonyls. The dicarbonyl augmented intracellular peroxides prior to the induction of HB-EGF mRNA as judged by flow cytometric analysis using 2',7'-dichlorofluorescin diacetate. N-acetyl-L-cysteine and aminoguanidine suppressed both dicarbonyl-increased HB-EGF mRNA and intracellular peroxide levels in RASMC. DL-Buthionine-(S,R)-sulfoximine increased the levels of 3-DG-induced HB-FGF mRNA. Furthermore, hydrogen peroxide alone also induced HB-EGF mRNA in RASMC. These results indicate that MG and 3-DG induce HB-EGF by increasing the intracellular peroxide levels. In addition, the pretreatment with 12-O-tetra-decanoylphorbol-13-acetate failed to alter dicarbonyl-induced HB-EGF mRNA expression in RASMC, suggesting that the signal transducing mechanism is not mediated by protein kinase C. Since HB-EGF is known as a potent mitogen for smooth muscle cells and is abundant in atherosclerotic plaques, the induction of HB-EGF by MG and 3-DG, as well as the concomitant increment of intracellular peroxides, may trigger atherogenesis during diabetes.

DOI PubMed CiNii

Molecular cloning and expression of amadoriase isoenzyme (fructosyl amine:oxygen oxidoreductase, EC 1.5.3) from Aspergillus fumigatus

M Takahashi, M Pischetsrieder, VM Monnier

JOURNAL OF BIOLOGICAL CHEMISTRY ( AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC )  272 ( 19 ) 12505 - 12507  1997年05月

　概要を見る

Amadoriase is an enzyme catalyzing the oxidative deglycation of Amadori products to yield corresponding amino acids, glucosone, and H2O2. We previously reported the purification and characterization of two amadoriase isozymes from Aspergillus sp, that degrade both glycated low molecular weight amines and amino acids (Takahashi, M., Pischetsrieder, M., and Monnier, V. M. (1997) J. Biol, Chem. 272, 3437-3443), To identify the primary structure of the enzymes, we have prepared a cDNA library from Aspergillus fumigatus induced with fructosyl propylamine and isolated a clone using polyclonal anti-amadoriase II antibody, The primary structure of the enzyme deduced from the nucleotide sequence comprises 438 amino acid residues with a predicted molecular mass of 48,798 Da. The deduced primary structure exhibits the presence of an ADP-binding motif near the NH2 terminus. The identity of the amadoriase II cDNA was further confirmed by expression in Escherichia coli cells with an inducible expression system, Northern blotting analysis revealed that amadoriase II was induced by fructosyl propylamine in a dose-dependent manner.

DOI PubMed CiNii

cDNA cloning of an amadoriase enzyme which cleaves glycated amino acids

M Takahashi, M Pischetsrieder, VM Monnier

INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE ( LIPPINCOTT-RAVEN PUBL )  38 ( 4 ) 5290 - 5290  1997年03月

研究発表ペーパー・要旨（国際会議）  

Isolation, purification, and characterization of amadoriase isoenzymes (fructosyl amine-oxygen oxidoreductase EC 1.5.3) from Aspergillus sp

M Takahashi, M Pischetsrieder, VM Monnier

JOURNAL OF BIOLOGICAL CHEMISTRY ( AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC )  272 ( 6 ) 3437 - 3443  1997年02月

　概要を見る

Four ''amadoriase'' enzyme fractions, which oxidatively degrade glycated low molecular weight amines and amino acids under formation of hydrogen peroxide and glucosone, were isolated from an Aspergillus sp. soil strain selected on fructosyl adamantanamine as sole carbon source. The enzymes were purified to homogeneity using a combination of ion exchange, hydroxyapatite, gel filtration, and Mono & column chromatography. Molecular masses of amadoriase enzymes Ia, Ib, and Ic were 51 kDa, and 49 kDa for amadoriase II. Apparent kinetic constants for N-epsilon-fructosyl N-alpha-t-butoxycarbonyl lysine and fructosyl adamantanamine were almost identical for enzymes Ia, Ib, and Ic, but corresponding values for enzyme II were significantly different. FAD was identified in all enzymes based on its typical absorption spectrum. N terminal sequence was identical for enzymes Ia and Ib (Ala-Pro-Ser-Ile-Leu-Ser-Thr-Glu-Ser-Ser-Ile-Ile-Val-Ile-Gly-Ala- Gly-Thr-Trp-Gly-) and Ic except that the first 5 amino acids were truncated. The sequence of enzyme II was different (Ala-Val-Thr-Lys-Ser-Ser-Ser-Leu-Leu-Ile-Val-Gly-Ala-Gly- Thr-Trp-Gly-Thr-Ser-Thr-). All enzymes had the FAD cofactor-binding consensus sequence Gly-X-Gly-X-X-Gly within the N-terminal sequence. In summary, these data show the presence of two distinct amadoriase enzymes in the Aspergillus sp. soil strain selected on fructosyl adamantanamine and induced by fructosyl propylamine. In contrast to previous described enzymes, these novel amadoriase enzymes can deglycate both glycated amines and amino acids.

DOI PubMed CiNii