A compound heterozygous mutation of the alkaline phosphatase ALPL gene causes hypophosphatasia in a Han Chinese family.
A compound heterozygous mutation of the alkaline phosphatase ALPL gene causes hypophosphatasia in a Han Chinese family.
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DOI：
10.3892/etm.2020.9281
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发表时间：
2020-12
影响因子：
2.7
通讯作者：
Xiong F
中科院分区：
文献类型：
--
作者：
Huang H;Wang J;Liang Y;Wei X;Guo D;Sun H;Zhang X;Xu X;Xiong F
磷酸酶减退症（HPP）是一种罕见的遗传性全身性疾病，以骨和/或牙齿矿化缺陷为特征，由碱性磷酸酶基因（ALPL）突变引起。本研究对中国汉族HPP家族的ALPL突变进行了研究，探讨了ALPL基因突变的发病机制。从家庭成员的外周静脉血中提取DNA。Sanger测序用于筛选突变。通过生物信息学、亚细胞定位、酶活性测定和western blotting分析了这两种突变的发病机制。Sanger测序显示复合杂合突变c.203C>T （p.T68M）和c.571G>A （p.E191K）。突变位于ALPL基因的外显子4和6，并通过polyphen2分析预测为有害突变。蛋白质分析表明，转染携带突变的质粒后，293T细胞的成熟蛋白产量下降，酶活性降低。将ALPL基因克隆到pcDNA3.1（+）载体上，构建突变质粒ALPL- pt68m和ALPL- pe191k。在转染ALPL-pE191K突变质粒的细胞中观察到的免疫荧光主要位于细胞膜。然而，与野生型相比，细胞质中的染色增加，转染ALPL-pT68M突变质粒的293T细胞几乎没有荧光。目前的研究结果表明，复合杂合的c.571G>A和c.203C>T突变可能通过导致定位错误、蛋白质表达降低和酶活性丧失而导致儿童HPP。本研究结果可能为HPP的潜在分子机制提供新的见解。
Hypophosphatasia (HPP) is a rare hereditary systemic disease that is characterized by defective bone and/or dental mineralization, and is caused by mutations in the alkaline phosphatase gene (ALPL). The present study investigated the ALPL mutation in a Chinese Han family with HPP and studied the pathogenesis of the mutations of the ALPL gene. DNA was extracted from peripheral venous blood of the family members. Sanger sequencing was used to screen the mutations. Associations between pathogenesis for both mutations were analyzed by bioinformatics, subcellular localization, measurement of enzyme activity and western blotting. Sanger sequencing revealed the compound heterozygous mutations c.203C>T (p.T68M) and c.571G>A (p.E191K). The mutations were located at exon 4 and 6 of the ALPL gene and were predicted by Polyphen-2 analysis to be harmful. Protein analysis indicated a decrease in mature protein production and lower enzyme activity in 293T cells transfected with plasmids carrying the mutations. The ALPL gene was cloned into the pcDNA3.1(+) vector and mutant plasmids ALPL-pT68M and ALPL-pE191K were constructed. Immunofluorescence observed in cells transfected with the ALPL-pE191K mutant plasmid was mainly located in the cell membrane. However, staining in the cytoplasm was increased compared with the wild type, and almost no fluorescence was identified in 293T cells transfected with the ALPL-pT68M mutant plasmid. The present findings demonstrated that the compound heterozygous c.571G>A and c.203C>T mutations may contribute to childhood HPP by resulting in mislocalization, decreased protein expression and loss of enzyme activity in a Han Chinese family. The results of the current study may provide insights into the potential molecular mechanism of HPP.
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影响因子：
13.8
作者：
Combet, C;Blanchet, C;Deléage, G
通讯作者：
Deléage, G
影响因子：
1.9
作者：
Mornet, Etienne;Yvard, Alice;Simon-Bouy, Brigitte
通讯作者：
Simon-Bouy, Brigitte
DOI：
10.1073/pnas.89.20.9924
发表时间：
1992-10-15
影响因子：
11.1
作者：
HENTHORN, PS;RADUCHA, M;WHYTE, MP
通讯作者：
WHYTE, MP
影响因子：
6.2
作者：
Orimo, H;Girschick, HJ;Shimada, T
通讯作者：
Shimada, T
影响因子：
4.8
作者：
Mornet, E;Stura, E;Le Du, MH
通讯作者：
Le Du, MH