摘要
真核细胞内多种无膜及有膜细胞器为各种生物学过程的发生提供场所.被膜细胞器通过它们之间的膜接触位点所进行的信息交流和物质交换是维持生命活动所必需的.绘制活细胞中细胞器或膜接触位点等处的蛋白质组图谱,将有助于解析这些部位的生物学功能及作用机制,并为研究细胞器相互作用提供基础.但由于无膜细胞器或膜接触位点很难分离纯化,传统的生化方法难以系统解析其中的蛋白质组.最近报道的几种基于酶类的蛋白质邻近标记技术,则为系统分析上述空间受限的蛋白质组这一难题提供了有效的解决方案.通过将能催化产生活性自由基(最常见的是生物素及其衍生物的自由基)的酶连接到目标蛋白上,可对其邻近的蛋白质组进行共价标记,从而使后者的分离和鉴定成为可能,并可以运用于活细胞中的动态标记.我们在此综述了几种最新的邻近标记策略的原理及应用,并对它们的优势与局限性进行了比较,以期为细胞器互作的蛋白质组学研究提供参考.
Membrane-less and membrane-bound organelles provide sites for many crucial biological processes to take place within eukaryotic cells.Signalling and exchange of materials occurring at membrane contact sites(MCSs)among membrane-bound organelles are also vital for cell homeostasis.Proteomic mapping of molecular machineries within membrane-less organelles or MCSs are essential for the understanding all those events within such sites,as well as for the study of organelle interactions.However,the attempts to dissect molecular determinants in these sites using traditional biochemical techniques were far from fruitful.Recent advances in proximity labelling techniques provide an ideal solution for this challenge.Mostly by utilizing different type of biotin ligases which fused to target proteins,proteomes surrounding the target protein(within tens of nanometers)could be tagged(often with biotin or its derivatives).These covalently labelled proteins could then be enriched and identified with mass spectrometry.Here,we reviewed most recent advances in proximity labeling approaches,hoping to provide a general guide for researches focusing on proteomic mapping of molecular machineries within organelles or MCSs.
作者
杜阳春
唐菁兰
王友军
张晓嫣
DU Yang-Chun;TANG Jing-Lan;WANG You-Jun;ZHANG Xiao-Yan(Ministry of Education Key Laboratory of Cell Proliferation and Regulation Biology,College of Life Sciences,Beijing Normal University,Beijing 100875,China;Beijing Key Laboratory of Gene Resource and Molecular Development,College of Life Sciences,Beijing Normal University,Beijing 100875,China;Tsinghua International School,Campus of Tsinghua High School,Beijing 100084,China)
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2019年第7期641-653,共13页
Progress In Biochemistry and Biophysics
基金
国家自然科学基金面上项目(31671492)
北京师范大学细胞增殖及调控生物学教育部重点实验室开放基金资助项目~~
