Research for properties of the expressed recombinant human serum cholinesterase variant

表达的重组人血清胆碱酯酶变异体的特性研究

基本信息

  • 批准号:
    08672651
  • 负责人:
  • 金额:
    $ 1.41万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
  • 财政年份:
    1996
  • 资助国家:
    日本
  • 起止时间:
    1996 至 1997
  • 项目状态:
    已结题

项目摘要

We have found individuals heterozygous for BCHE L330I from a low serum BCHE activity group and slightly low DN and FN group selected randomly from the population. A 63-year-old Japanese man showed low serum BCHE activity on health examination. Secondary hypocholinesterasemia due to agricultural chemical poisoming and severe hepatic dysfunction were ruled out. The phenotyping analysis revealed a reduced dibucaine number (DN) and an esxpecially low fluoride number (FN), similar to an FS phenotype. Although the fluoride-resistant gene has been reported to be caused by T243M and G390V in Caucasian, we could not find these mutations in Japanese containing this case. A homozygousmissense mutation, a T to A transversion at nucleotide 988, was identified in his BCHE gene. This mutation resulted in the replacement of leusine by isoleucine at codon 330 (L339I).It is important to demonstrate whether the abnormal DN and FN are originated from the BCHE L330I mutation alone or there are some other genetic or modifying factors. DN and FN of recombinant BCHE (L339I) secreted by human fetal kidney cells were compared to recombinant wild-type (usual gene) BCHE and normal serum BCHE.The individual nucleotide substitution characterizing the BCHE L330I variant was introduced into the pRcCMV plasmid containing usual BCHE using PCR.By expression of the recombinant BCHE protein in human fetal kidney cells (293 ; JCRB9068、ATCC CRL1573) which were transformed with purified pRcCMV-BCHE plasmid, we showed that the BCHE L330I variant resulted in low enzymatic activity and low DN and FN.These results showed this amino acid substitution of BCHE,Leu330 to Ile, really caused the abnormal DN and FN.We conclude the BCHE L330I mutation is a fluoride-resistant gene, a Japanese type fluoride-resistant gene.
我们从人群中随机抽取低血清BCHE活性组和轻度低DN、FN组,发现BCHE L330 I杂合子。一位63岁的日本男性在健康检查时显示出低血清BCHE活性。排除了农药中毒和严重肝功能障碍引起的继发性低胆碱酯酶血症。表型分析显示,减少的地布卡因数(DN)和特别是低氟值(FN),类似于FS表型。虽然在白种人中已报道耐氟基因是由T243 M和G390 V引起的,但我们在包含该病例的日本人中未发现这些突变。在该患者的BCHE基因中发现了一个同源错义突变,即988位核苷酸的T → A颠换。该突变导致第330位亮氨酸被异亮氨酸取代(L339 I),提示DN和FN的异常是由BCHE L330 I突变引起的,还是存在其他遗传或修饰因素。将人胎肾细胞分泌的重组BCHE(L339 I)的DN和FN与重组野生型进行比较,将BCHE L330 I变异体的核苷酸序列插入到含有BCHE L330 I变异体的pRcCMV质粒中,在人胚肾细胞中表达重组BCHE蛋白,(293 ; JCRB 9068,ATCC CRL 1573)中的BCHE突变,结果表明BCHE L330 I突变导致了低酶活和低DN、FN,说明BCHE L330 I突变是导致DN、FN异常的真正原因,我们认为BCHE L330 I突变是一个耐氟基因,一种日本型抗氟基因。

项目成果

期刊论文数量(4)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Kayoko Sudo: "Human butyrylcholinesterase mutation L330I belong to a fluorid-resistant gene,by expression in human fetal kidney cells" Biochem Biophys Res Comm. 240. 312-375 (1997)
Kayoko Sudo:“人丁酰胆碱酯酶突变 L330I 属于耐氟基因,在人胎儿肾细胞中表达”Biochem Biophys Res Comm。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
Dey DC,Maekawa M,sudo K,Kanno T.: "Butyrylcholinesterase genes in individuals with abnormal inhibition numbers and with trace activity : one common mutation and two movel silent genes." Ann Clin Biochem. (in press).
Dey DC、Maekawa M、sudo K、Kanno T.:“抑制数量异常且具有微量活性的个体中的丁酰胆碱酯酶基因:一种常见突变和两种 movel 沉默基因。”
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
Kayoko Sudo: "Human butyrylcholinesterase mutation L330I belong to a fluoridresistant gene,by expression in human fetal kidney cells" Biochem Biophys Res Comm. 240. 372-375 (1997)
Kayoko Sudo:“人丁酰胆碱酯酶突变 L330I 属于耐氟基因,在人胎儿肾细胞中表达”Biochem Biophys Res Comm。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
M.Maekawa: "Genetic mutations of butyrylcholinesterase identified from its phenotypic abnormalities in Japan." Clinical Chemistry. (in press).
M.Maekawa:“从日本的表型异常中发现了丁酰胆碱酯酶的基因突变。”
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SUDO Kayoko其他文献

SUDO Kayoko的其他文献

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{{ truncateString('SUDO Kayoko', 18)}}的其他基金

BIOCHEMICAL PROPERTIES OF RECOMBINANT CHOLONESTERASE VARIANT SPECIFIC IN JAPANESE
日本特有的重组胆酯酶变体的生化特性
  • 批准号:
    10672187
  • 财政年份:
    1998
  • 资助金额:
    $ 1.41万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Genetic Analysis of Lactate Dehydrogenase (LDH)-H(B) Subunit Variant
乳酸脱氢酶(LDH)-H(B)亚基变异体的遗传分析
  • 批准号:
    03671116
  • 财政年份:
    1991
  • 资助金额:
    $ 1.41万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (C)
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