High-throughput experimental methods for investigating biomolecular condensates

查看全文 作　　者：Taoyu [1]Chen;Qi [1]Lei;Minglei [2]Shi;Tingting [1]Li 机构地区：[1]Department of Biomedical Informatics,School of Basic Medical Sciences,Peking University Health Science Center,Beijing 100191,China;[2]MOE Key Laboratory of Bioinformatics,Bioinformatics Division and Center for Synthetic&Systems Biology,TNLIST,School of Medicine,Tsinghua University,Beijing 100084,China 出　　处：《Quantitative Biology》索引2021年第9卷第3期,共12页 基　　金：This work was supported by the National Key Research and Development Program of China(2018YFA0507504);the National Natural Science Foundation of China(61773025,32070666);Clinical Medicine Plus X-Young Scholars Project of Peking University(PKU2021LCXQO12);the Fundamental Research Funds for the Central Universities. 摘　　要：Background:The concept of biomolecular condensate was put forward recently to emphasize the ability of certain cellular compartments to concentrate molecules and comprise proteins and nucleic acids with specific biological functions,from ribosome genesis to RNA splicing.Due to their unique role in biological processes,it is crucial to investigate their compositions,which is a primary determinant of condensate properties.Results:Since a wide range of macromolecules comprise biomolecular condensates,it is necessary for researchers to investigate them using high-throughput methodologies while low-throughput experiments are not efficient enough.These high-throughput methods usually purify interacting protein libraries from condensates before being scanned in mass spectrometry.It is possible to extract organelles as a whole for specific condensates for further analysis,however,most condensates do not have a distinguishable marker or are sensitive to shear force to be extracted as a whole.Affinity tagging allows a comprehensive view of interacting proteins of target molecule yet only proteins with strong bonds may be pulled down.Proximity labeling serves as a complementary method to label more dynamic proteins with weaker interactions,increasing sensitivity while decreasing specificity.Image-based fluorescent screening takes another path by scanning images automatically to illustrate the condensing state of biomolecules within membraneless organelles,which is a unique feature unlike the previous mass spectrometry-based methods.Conclusion:This review presents a rough glimpse into high-throughput methodologies for biomolecular condensate investigation to encourage usage of bioinformatic tools by researchers in relevant fields. 关 键 词：biomolecular condensates HIGH-THROUGHPUT phase separation INTERACTION