S. Takegawa
University of Tokyo
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British Journal of Haematology | 1980
Shiro Miwa; Hisaichi Fujii; S. Takegawa; T. Nakatsuji; Kunio Yamato; Yoji Ishida; N. Ninomiya
Summary Seven new red‐cell pyruvate kinase (PK) variants were characterized by the methods recently recommended by the International Committee for Standardization in Haematology. The cases were all true homozygote as evidenced by consanguineous marriages of the parents; all are Japanese. These variants were designated as PK Tokyo, PK Nagasaki, PK Sapporo, PK Maebashi, PK Itabashi, PK Fukushima and PK Aizu, respectively. Low substrate affinity (high K0.5S for phosphoenolpyruvate) and thermal instability appear to play major roles in causing defective enzyme function, resulting in chronic haemolytic anaemia. Product inhibition of PK by ATP may also play an additional role in causing haemolysis in more than half the cases.
British Journal of Haematology | 1983
S. Takegawa; Hisaichi Fujii; Shiro Miwa
Summary. The relationship between erythroid cell maturation and the change of pyruvate kinase (PK) isozymes was studied using fluorescent antibody techniques. The fluorescence of M2‐type PK was strongest in the proerythroblast stage, and then steeply declined with red cell maturation so that only weak fluorescence was seen in orthochromatic erythroblasts. On the other hand, the fluorescence of L‐type PK was seen throughout the maturation of the erythroblasts, with stronger fluorescence in the basophilic or polychromatic erythroblasts than in proerythroblasts.
Human Genetics | 1981
Hisaichi Fujii; Shiro Miwa; Kenzaburo Tani; S. Takegawa; Noboru Fujinami; Kenji Takahashi; Shin Nakayama; M. Konno; Tetsuya Sato
SummaryTwo new glucose 6-phosphate dehydrogenase (G6PD) variants associated with chronic nonspherocytic hemolytic anemia were discovered. G6PD Kobe was found in a 16-year-old male associated with hemolytic crisis after upper respiratory infection. The enzyme activity of the variant was about 22% of that of the normal enzyme. The main enzymatic characteristics were slower than normal anodal electrophoretic mobility, high Km G6P, increased thermal-instability, an acidic pH optimum, and an extremely increased affinity for the substrate analogue, galactose 6-phosphate (Gal-6P).G6PD Sapporo was found in a 3-year-old male associated with drug-induced hemolysis. The enzyme activity was extremely low, being 3.6% of normal. In addition, this variant showed high Ki NADPH and thermal-instability.G6PD Kobe utilized the artificial substrate Gal-6P effectively as compared with the common natural substrate, glucose 6-phosphate. In G6PD Sapporo, NADPH could not exert the effect of product inhibition. The structural changes of these variants are expected to occur at the portions inducing conformational changes of the substrate binding site of the enzyme.
Human Genetics | 1984
T. Takizawa; Hisaichi Fujii; S. Takegawa; Kenji Takahashi; Akira Hirono; Takayuki Morisaki; Hitoshi Kanno; R. Oka; H. Yoshioka; Shiro Miwa
SummaryA new glucose 6-phosphate dehydrogenase (G6PD) variant associated with chronic nonspherocytic hemolytic anemia was discovered in Japan. The patient showed hemolytic crises after upper respiratory infections. The enzyme activity was about 3.8% of the normal. The partially purified enzyme revealed slow anodal electrophoretic mobility, high Km NADP, marked thermal-instability, and increased affinity for a substrate analogue (deamino-NADP). A particular characteristic of this enzyme was a biphasic pH curve with a greatly increased activity at low pH values. From these results, this variant was clearly different from hitherto observed G6PD variants, and was designated G6PD Asahikawa.
Human Genetics | 1983
Takayuki Morisaki; Hisaichi Fujii; S. Takegawa; Kenzaburo Tani; Akira Hirono; T. Takizawa; Kenji Takahashi; M. Shinogi; T. Teshirogi; Shiro Miwa
SummaryA new glucose-6-phosphate dehydrogenase (G6PD) variant associated with chronic nonspherocytic hemolytic anemia was discovered. It was found in a 2-year-old male who had a hemolytic crisis after an upper respiratory tract infection. The enzyme activity of the variant was 8.4% of that of the normal enzyme. The enzymatic characteristics were slower than normal anodal electrophoretic mobility, low Km G6P, normal Km NADP, increased utilization of substrate analogues, high Ki NADPH, decreased heat stability, and an alkaline pH optimum. From these results, this was considered to be a new variant and was designated G6PD Sendagi.
Human Genetics | 1984
Hisaichi Fujii; Shiro Miwa; S. Takegawa; Kenji Takahashi; Akira Hirono; T. Takizawa; Takayuki Morisaki; Hitoshi Kanno; T. Taguchi; J. Okamura
SummaryTwo new glucose-6-phosphate dehydrogenase (G6PD) variants were discovered in Japan. The first, found in a 9-year-old male, was associated with chronic hemolysis and hemolytic crises after upper respiratory infections. The enzyme activity of the variant was 2.9% of normal. The patients G6PD showed an increased utilization of substrate analogue, deamino-NADP, and thermal instability. The second variant occurred in a 7-year-old male with druginduced hemolysis. The main enzymatic characteristics were reduced enzyme activity, being 6.4% of normal, faster-thannormal anodal electrophoretic mobility, slightly high Michaelis constant for glucose-6-phosphate, thermal instability, and biphasic pH optima. Enzymatic properties of these variants allowed each to be distinguished from previously reported variants. The first variant was designated Gd (-) Gifu and the other, Gd (-) Fukuoka.
Vox Sanguinis | 1981
Yoji Ishida; Shiro Miwa; S. Takegawa; Noboru Fujinami; Kunio Yamato
Abstract. Red cells from 5 healthy adults and an unstable mutant pyruvate kinase (PK), PK Maebashi, were stored for 12 months in a ‐80°C freezer. With an exception of triosephosphate isomerase and phosphoglycerate kinase activities, normal red‐cell enzyme activities remained essentially unchanged. Mutant PK was characterized by the methods recommended by the International Committee for Standardization in Haematology (ICSH). Normal control and PK Maebashi remained essentially unchanged in their characteristics after storage in a ‐80°C freezer for 12 months. These results indicate that the red cells stored over a long term in a ‐80°C freezer can be used for the study of unstable mutant PK.
American Journal of Hematology | 1981
Shiro Miwa; Hisaichi Fujii; Kenzaburo Tani; Keisuke Takahashi; S. Takegawa; Noboru Fujinami; Minoru Sakurai; Masako Kubo; Yasuo Tanimoto; Tomoaki Kato; Noboru Matsumoto
American Journal of Hematology | 1981
Shiro Miwa; Yoji Ishida; S. Takegawa; Gumpei Urata; Toshio Toyoda
American Journal of Hematology | 1981
Yoji Ishida; Shiro Miwa; Hisaichi Fujii; Noboru Fujinami; S. Takegawa; Kunio Yamato