Erythroenzymopathies: Mechanisms of isozyme switches and clarification of genetic abnormalities

红酶病：同工酶转换机制和遗传异常的澄清

• 批准号: 60480279
• 负责人: MIWA Shiro
• 金额: $ 4.29万
• 依托单位: University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1985
• 资助国家: 日本
• 起止时间: 1985 至 1986
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-60480279/
• 关键词: hereditary hemolytic anemia; erythroenzymopathies; pyruvate kinase deficiency; human L-type PK cDNA; L型PK遺伝子座

We discovered 7 new cases of glucose 6-phosphate dehydrogenase deficiency and 3 new cased of pyruvate kinase (PK) deficiency after the survey of patients showing non-spherocytic hemolytic anemia due to unknown causes. In addition to these works, we could clone the cDNA for liver-type PK and assign its chromosomal locus as a first step for the clarification of molecular anomalies. PK has four isozymes (L, R, <M_1> , <M_2> ) that are encoded mainly by two different genes. We isolated a cDNA clone from a Japanese adult liver <lambda> gt10 cDNA library by using a rat liver (L)-type PK cDNA probe. One positively hybridizing clone, hlPK-1, which contained a 1,049-base pair cDNA insert, was subjected to DNA sequence analysis.Comparisons of the sequence data with the rat PK cDNAs indicated that the cDNA encoded information for the carboxyl terminal 105 amino acids of a human L-type PK and a 3'untranslated region of 734 nucleotides. Furthermore, the karyotype analysis of several human-mouse hybrid cells and Southern blot analysis of DNAs of the hybrids with a hlPK-1 indicated that the human L-type PK gene is located on chromosome 1.

我们发现7例新的葡萄糖6-磷酸脱氢酶缺乏症和3例新的丙酮酸激酶（PK）缺乏症后，患者的调查显示非球形细胞溶血性贫血，由于不明原因。除了这些工作，我们可以克隆肝型PK的cDNA，并指定其染色体位点作为澄清分子异常的第一步。PK有四种同工酶（L，R，<M_1>，<M_2>），主要由两个不同的基因编码。我们<lambda>用大鼠肝（L）型PK cDNA探针从日本成人肝gt 10 cDNA文库中分离了一个cDNA克隆。对一个含有1，049个碱基的cDNA插入片段的阳性杂交克隆hlPK-1进行DNA序列分析，与大鼠PK cDNA序列比较表明，该cDNA编码人L型PK羧基端105个氨基酸和3 '端734个核苷酸的非翻译区。此外，几种人-小鼠杂交细胞的核型分析和具有hlPK-1的杂交细胞的DNA的Southern印迹分析表明，人L型PK基因位于1号染色体上。

项目成果

Morisaki T;Horiuchi N;Fujii H;Miwa S: Amer.J.Hematol.23. 263-269 (1986)

Morisaki T；Horiuchi N；Fujii H；Miwa S：Amer.J.Hematol.23。

Hirono A;Fujii H;Natori H;Kurokawa I;Miwa S: Brit.J.Haemat.

Hirono A；Fujii H；Natori H；Kurokawa I；Miwa S：Brit.J.Haemat。

Hirono, A., Fujii, H., Natori, H., Kurokawa, I., and Miwa, S.: "Chromatographic analysis of human erythrocyte pyrimidine 5'-nucleotidase from five patients with pyrimidine 5'-nucleotidase deficiency" Brit. J. Haemat.in press.

Hirono, A.、Fujii, H.、Natori, H.、Kurokawa, I. 和 Miwa, S.：“五名嘧啶 5-核苷酸酶缺乏症患者的人红细胞嘧啶 5-核苷酸酶的色谱分析”英国。

Miwa S;Fujii H;Tani K;Miyamoto T;Nishida Y: Jpn.J.Exp.Med.56. 293-296 (1986)

Miwa S；Fujii H；Tani K；Miyamoto T；Nishida Y：Jpn.J.Exp.Med.56。

Tani, K., Fujii, H., Tsutsumi, H., Sukegawa, J., Toyoshima, K., Yoshida, M-C., Noguchi, T., Tanaka, T., and Miwa, S.: "Human liver type pyruvate kinase: cDNA cloning and chromosomal assignment" Biochem. Biophys. Res. Commun.in press.

Tani, K.、Fujii, H.、Ttsutsumi, H.、Sukekawa, J.、Toyoshima, K.、Yoshida, M-C.、Noguchi, T.、Tanaka, T. 和 Miwa, S.：“人类肝脏类型