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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

基本信息

批准号：
10672187
负责人：
SUDO Kayoko
金额：
$ 1.98万
依托单位：
KOKUSAI GAKUIN SAITAMA JUNIOR COLLAGE (2000)Jikei University School of Medicine (1998-1999)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10672187/
关键词：
SERUM CHOLINESTERASE VARIANT GENE GENE EXPRESSION MISSENSE MUTATION 293 CELL EXPRESSION VECTOR FLUORIDE-RESISTANT ONE POINT MUTATION フカソーダ耐性

项目摘要

Several genetic variants of human serum butyrylcholinesterase (EC3.1.1.8 ; BCHE) have been reported to be associated with prolonged apnea after medication of the muscle relaxant drug succinylcholine. These BCHE variants were qualitatively identified by dibucaine number (DN) and fluoride number (FN). Atypical variant is phenotyped by resistance to inhibition by dibucaine (low DN) and fluoride resistant variants were phenotyped by resistance to inhibition by NaF (low FN and DN).Two types of fluoride-resistant gene have been reported. They were Fluoride-1 and -2 variants which were Thr243Met and Gly390Val, respectively. Even after screening several times for the atypical BCHE (Asp70Gly), and two kinds of the fluoride-resistant variants in the Japanese population, we could not find these mutations (our personal data). Therefore, we considered these differences might have originated from ethnic difference.We found an individual homozygous for L330I mutation (alteration from leucine to isoleucine at codon 330) who showed reductions of enzymatic activity, DN and FN.His DN and FN showed FS phenotype rather than FF.The L330I mutation has never been reported in foreign countries.By expression of the recombinant BCHE protein in human fetal kidney cells, The BCHE L330I variant showed low enzymatic activity, low DN and FN.Therefore, we conclude the BCHE L330I mutation is a Japanese type fluoride-resistant gene. We also have expressed recombinant ChE proteins (wild type, 330V and 330S). All of the recombinants with amino acid substitutions at codon 330 caused not only a reduction of BCHE activity but also the abnormal DN and FN.The degrees of the reduction were almost the same. We conclude that the mutant recombinant BCHEs at codon 330 possibly have the properties of the fluoride-resistant gene and suggest that codon 330 is important to interact with substrate and sodium fluoride.

据报道，人血清丁酰胆碱酯酶（EC3.1.1.8 ; BCHE）的几种遗传变异与肌肉松弛药物琥珀酰胆碱给药后的长时间呼吸暂停有关。通过地布卡因值（DN）和氟化物值（FN）对这些BCHE变体进行定性鉴定。非典型变异体的表型是对地布卡因抑制的抗性（低DN），而耐氟变异体的表型是对NaF抑制的抗性（低FN和DN）。它们是氟化物-1和氟化物-2变异体，分别为Thr 243 Met和Gly 390 Val。即使在对日本人群中的非典型BCHE（Asp 70 Gly）和两种耐氟变体进行多次筛查后，我们也无法发现这些突变（我们的个人数据）。因此，我们认为这些差异可能源于种族差异，我们发现了一个L330 I突变的纯合子个体，（在密码子330处从亮氨酸变为异亮氨酸）的患者显示酶活性降低，结果表明，BCHE基因在人胎肾中表达，其L330 I突变在国外未见报道BCHE L330 I突变体在细胞中表现出低酶活性、低DN和低FN，因此我们认为BCHE L330 I突变体是一个日本型耐氟基因。我们还表达了重组ChE蛋白（野生型，330 V和330 S）。所有第330位密码子发生氨基酸替换的重组子不仅引起BCHE活性降低，而且引起DN和FN异常，其降低程度基本相同。我们的结论是，在密码子330的突变重组BCHE可能具有耐氟基因的性质，并建议密码子330是重要的与底物和氟化钠的相互作用。