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引用本文:   王舒越, 贾胜男, 李潇影, 张海洋, 冯馨, 王诗, 陶纬国, 胡良海. 人尿液外泌体蛋白质组的糖基化修饰分析. 分析化学, 2020, 48(10): 1334-1342. doi:  10.19756/j.issn.0253-3820.201220 [复制]

Citation:   WANG Shu-Yue , JIA Sheng-Nan , LI Xiao-Ying , ZHANG Hai-Yang , FENG Xin , WANG Shi , TAO Wei-Guo , HU Liang-Hai . A Preliminary Study of Human Urine Exosome Glycoproteome. Chinese Journal of Analytical Chemistry, 2020, 48(10): 1334-1342. doi: 10.19756/j.issn.0253-3820.201220 [复制]

人尿液外泌体蛋白质组的糖基化修饰分析

通讯作者:  胡良海, lianghaihu@jlu.edu.cn

收稿日期: 2020-04-21

基金项目: 本文系国家自然科学基金项目(No.21675061)资助

A Preliminary Study of Human Urine Exosome Glycoproteome

Corresponding author:  HU Liang-Hai , lianghaihu@jlu.edu.cn

Received Date:  2020-04-21

Fund Project:  This work was supported by the National Natural Science Foundation of China (Nos. 21675061).

人体尿液外泌体糖基化修饰可反映多种生物信息,发展规模化的鉴定方法对深入研究其功能至关重要。本研究针对尿液外泌体糖基化修饰提取富集的关键问题,发展了一种结合EVTRAP磁珠(Extracellular vesicles total recovery and purification)和点击麦芽糖材料两步提取富集的研究策略。基于外泌体的磷脂双分子层的特点,利用亲脂性EVTRAP磁珠从30 mL尿液中有效提取外泌体,经裂解和酶解后,再基于糖肽糖链部分的亲水性特点,结合同样具有亲水性的点击麦芽糖材料对蛋白酶解液中的糖肽进行有效富集,利用液相色谱-串联质谱联用(LC-MS-MS)技术对尿液外泌体全蛋白和糖基化修饰进行检测鉴定。最终鉴定到1925种非冗余尿液外泌体蛋白质和14233条肽段,实现了对尿液外泌体的有效提取;同时鉴定到来自88种糖蛋白的468条非冗余的糖肽,对应修饰于135个位点上的95种不同糖链组成,实现了对尿液外泌体糖基化蛋白质组的规模化鉴定,并从糖蛋白修饰异质性和糖基化类型的角度进行整理分析,为尿液外泌体糖基化修饰的研究提供了更有效的分析方法。

关键词:   外泌体, 糖基化, 尿液, EVTRAP磁珠, 点击麦芽糖
Key words:   Exosome, Glycosylation, Urine, Extracellular vesicles total recovery and purification, Click maltose
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人尿液外泌体蛋白质组的糖基化修饰分析

王舒越, 贾胜男, 李潇影, 张海洋, 冯馨, 王诗, 陶纬国, 胡良海

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