上海交通大学学报（医学版）

上海交通大学学报(医学版) ›› 2020, Vol. 40 ›› Issue (12): 1571-1578.doi: 10.3969/j.issn.1674-8115.2020.12.001

• 论著·基础研究 • 上一篇    下一篇

葡萄糖 -6- 磷酸脱氢酶基因4种突变及蛋白结构分析

朱之星1, 2，纪 伟3，顾坚磊1, 4，吕 晖1, 2, 4，田国力3   

  1. 1. 上海交通大学附属儿童医院，上海市儿童医院生物医学信息研究中心，上海 200040；2. 上海交通大学附属儿童医院儿童精准医学大数据工程技术研究中心，上海 200040；3. 上海交通大学附属儿童医院新生儿筛查中心，上海 200040；4. 上海交通大学生命科学技术学院，上海交通大学－耶鲁大学生物统计与数据科学联合中心，上海 200240
  • 出版日期:2020-12-28 发布日期:2021-02-02
  • 通讯作者: 田国力，电子信箱：Tiangl@shchildren.com.cn。
  • 作者简介:朱之星（1996—），女，硕士生，电子信箱：981164372@qq.com。
  • 基金资助:
    国家重点研发计划（2018YFC0910500）；上海市科学技术委员会专项基金（17DZ2251200）；上海市卫生与计划生育委员会研究课题（2018ZHYL0223）；上海交通大学医工交叉项目（ZH2018QNA30）；上海市科学技术委员会科研项目（18441905100）；上海市重中之重临床重点专科建设项目（2017ZZ02019）。

Analysis of four mutations and protein structure of glucose-6-phosphate dehydrogenase gene

ZHU Zhi-xing1, 2, JI Wei3, GU Jian-lei1, 4, LÜ Hui1, 2, 4, TIAN Guo-li3   

  1. 1. Center for Biomedical Informatics, Shanghai Children’s Hospital, Shanghai Jiao Tong University, Shanghai 200040, China; 2. Shanghai Engineering Research Center for Big Data in Pediatric Precision Medicine, Shanghai 200040, China; 3 Newborn Screening Center, Shanghai Children’s Hospital, Shanghai 200040, China; 4. SJTU-Yale Joint Center for Biostatistics, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China
  • Online:2020-12-28 Published:2021-02-02
  • Supported by:
    National Key R&D Program of China (2018YFC0910500); Foundation of Science and Technology Commission of Shanghai Municipality (17DZ2251200); Foundation of Shanghai Municipal Commission of Health and Family Planning (2018ZHYL0223); Medical Conversion Cross Fund of Shanghai Jiao Tong University (ZH2018QNA30); Action Plan for Scientific and Technological Innovation in Shanghai (18441905100); Key Disciplines of Top Priority in Shanghai (2017ZZ02019).

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摘要： 目的·研究葡萄糖-6-磷酸脱氢酶（glucose-6-phosphate dehydrogenase，G6PD）基因突变对酶蛋白结构及功能的影响，了解G6PD突变与酶学结构、功能的关系。方法·回顾性分析2014—2017年上海市儿童医院205 103例新生儿G6PD缺乏症筛查记录，Wilcoxon 检验分析酶活性与突变基因类型之间的关系。采用Psipred、SOPMA、JPred4预测蛋白的二级结构，SWISS-MODEL 预测氨基酸链空间结构，Mupro、SDM、CUPSAT、mSCM、DUET、Dynamut预测蛋白突变前后稳定性变化，PROVEAN预测对蛋白功能的影响，PyMOL 和LigPlot+修饰蛋白空间结构预测图。结果·230例样本酶活性检测阳性，对其中121例进行基因检测，共检出8种突变，除c.95A>G、c.1376G>T、c.1388G>A 3种常见突变，c.1024C>T也表现出高发生率。4种突变均导致蛋白结构改变，稳定性降低，对功能有不良影响；此外，c.1388G>A突变导致其远离甘油配体结合区。结论·4种突变均通过改变蛋白结构进而降低蛋白的稳定性，其中c.1388G>A突变还会影响蛋白与底物的结合能力。

关键词: 葡萄糖-6-磷酸脱氢酶(G6PD)缺乏症, 基因突变, 酶活性, 酶蛋白结构, 蛋白稳定性

Abstract:

Objective · To analyze the effect of glucose-6-phosphate dehydrogenase (G6PD) gene mutations on the structure and function of enzymes. Methods · A retrospective analysis of G6PD screening records of 205 103 neonates collected in the Shanghai Children's Hospital from 2014 to 2017 was performed. Wilcoxon test was used to analyze the enzyme activity and mutant genes. The software tools Psipred, SOPMA, and JPred4 were used to predict the secondary structure of the protein, and the 3D structure of G6PD was predicted based on the amino acid chain by SWISS-MODEL and modified by PyMOL and LigPlot+. Six different analysis software programs, including Mupro, SDM, CUPSAT, mSCM, DUET, and Dynamut, were utilized to compare protein stability of the wild-type with the mutant forms. PROVEAN was used to analyze the effect of amino acid changes on the enzyme. Results · Two hundred and thirty samples were positive for enzyme deficiency, of which 121 positive samples were genetically tested and eight mutations were identified. Three common mutations c.95A>G, c.1376G>T, and c.1388G>A were present, and c.1024C>T mutation was identified in 14 samples. These four mutations induced a change in protein structure, reduced protein stability, and had adverse effects on function. In addition, c.1388G was adjacent to the glycerol ligand binding region in the wild type structure, whereas the c.1388G>A mutation caused this residue further away from it. Conclusion · Four nonsynonymous mutations (c.95A>G, c.1376G>T, c.1388G>A and c.1024C>T) reduce the stability of the G6PD enzyme by changing the structure of the protein, in which the c.1388G>A mutation also affects the binding ability of the protein to the substrate.

Key words: glucose-6-phosphate dehydrogenase (G6PD) deficiency, gene mutation, enzyme activity, protein structure, protein stability

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