亲电脂质分子修饰位点的定量化学蛋白质组学研究
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- 英文论文题名：Quantitative chemoproteomic site-mapping of protein modifications by lipid-derived electrophiles
- 论文作者：陈影 ; 王初
- 英文论文作者：Ying Chen ; Chu Wang ; Department of Chemistry ; Peking University ; Peking-Tsinghua Center for Life Science ; Peking University
- 年：2016
- 作者机构：北京大学化学与分子工程学院北京大学;北京大学-清华大学生命科学联合中心;
- 论文关键词：LDE ; 化学蛋白质组学
- 会议召开时间：2016-07-01
- 会议录名称：中国化学会第30届学术年会摘要集-第二十八分会：化学生物学
- 语种：中文
- 分类号：O629.73
- 学会代码：ZGHY
- 会议名称：中国化学会第30届学术年会-第二十八分会 ; 化学生物学
- 会议地点：中国辽宁大连
- 主办单位：中国化学会
- 学会名称：中国化学会
- 页数：1
- 文件大小：307k
- 原文格式：D
- 会议级别：全国
摘要
细胞膜性结构富含不饱和脂肪酸,如油酸(linoleic acid,LA)、花生四烯酸(arachidonic acid,AA),当细胞处于氧化应激条件时,活性氧自由基(reactive oxygen species,ROS)会攻击这些不饱和脂肪酸,从而降解生成一系列具有高度亲电性的物种,诸如:丙烯醛(acrolein,ACR)、4羟基壬烯醛(4-hydroxynonenal,HNE)。脂源性的亲电小分子(lipid-derived electrophile,LDE)容易与蛋白质和核酸上的亲核性残基进行迈克尔加成反应,尤其是蛋白质中的半胱氨酸,从而造成其活性的改变以及功能的丧失。在这个课题中,我们针对LDE分子,设计了高反应活性的化学探针,通过生物正交反应引入同位素标记的可切割的标签,经过后续的生物素富集,胰酶酶切以及位点切割,我们可以对1000多个半胱氨酸进行鉴定和定量。该方法可以应用于不同种类的LDE分子修饰靶点的鉴定,具有普适性。
Biological membranes are rich in polyunsaturated fatty acids such as linoleic acid(LA) and arachidonic acid(AA).When cells are under oxidative stress,reactive oxygen species(ROS) can attack these membrane lipids to break them down into a series of highly reactive electrophilic products,such as acrolein(ACR)、4- hydroxynonenal(HNE).These lipid-derived electrophile(LDE) molecules are able to react with nucleophilic residues in both DNA and proteins to form covalent adducts,thus imposing great impact on their structures and functions.In this work,we designed a chemical probe to react with LDEs modified proteins.Following ligation to an isotopic cleavable biotin tag,trypsin digestion and peptides release,we are able to identify and quantify over 1000 residue sites modified by LDEs in the human cellular proteomes.
Biological membranes are rich in polyunsaturated fatty acids such as linoleic acid(LA) and arachidonic acid(AA).When cells are under oxidative stress,reactive oxygen species(ROS) can attack these membrane lipids to break them down into a series of highly reactive electrophilic products,such as acrolein(ACR)、4- hydroxynonenal(HNE).These lipid-derived electrophile(LDE) molecules are able to react with nucleophilic residues in both DNA and proteins to form covalent adducts,thus imposing great impact on their structures and functions.In this work,we designed a chemical probe to react with LDEs modified proteins.Following ligation to an isotopic cleavable biotin tag,trypsin digestion and peptides release,we are able to identify and quantify over 1000 residue sites modified by LDEs in the human cellular proteomes.
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