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Chinese Academy of Sciences Institutional Repositories Grid
Sensitive profiling of cell surface proteome by using an optimized biotinylation method

文献类型:期刊论文

作者Li, Yanan1,2; Wan, Yan1,2; Mao, Jiawei1,2; Yao, Yating1,2; Wang, Keyun1; Qiao, Qinglong1; Fang, Zheng1,2; Ye, Mingliang1
刊名Journal of proteomics
出版日期2019-03-30
卷号196页码:33-41
ISSN号1874-3919
关键词Cell surface proteins Biotinylation Plasma membrane proteins Proteomics Breast cancer cell lines
DOI10.1016/j.jprot.2019.01.015
通讯作者Ye, mingliang(mingliang@dicp.ac.cn)
英文摘要Cell surface proteins are responsible for many critical functions. systematical profiling of these proteins would provide a unique molecular fingerprint to classify cells and provide important information to guide immunotherapy. cell surface biotinylation method is one of the effective methods for cell surface proteome profiling. however, classical workflows suffer the disadvantage of poor sensitivity. in this work, we presented an optimized protocol which enabled identification of more cell surface proteins from a smaller number of cells. when this protocol was combined with a tip based fractionation scheme, 4510 proteins, including 2055 annotated cell surface-associated proteins, were identified with only 20 microgram protein digest, showing the superior sensitivity of the approach. to enable process 10 times fewer cells, a pipet tip based protocol was developed, which led to the identification of about 600 cell surface-associated proteins. finally, the new protocol was applied to compare the cell surface proteomes of two breast cancer cell lines, bt474 and mcf7. it was found that many cell surface-associated proteins were differentially expressed. the new protocols were demonstrated to be easy to perform, time-saving, and yielding good selectivity and high sensitivity. we expect this protocol would have broad applications in the future. significance: cell surface proteins confer specific cellular functions and are easily accessible. they are often used as drug targets and potential biomarkers for prognostic or diagnostic purposes. thus, efficient methods for profiling cell surface proteins are highly demanded. cell surface biotinylation method is one of the effective methods for cell surface proteome profiling. however, classical workflows suffer the disadvantage of poor sensitivity. in this work, we presented an optimized protocol which enabled identification of more cell surface proteins from a smaller number of starting cells. the new protocol is easier to perform, time-saving and has less protein loss. by using a special pipet tip, sensitive and in-depth cell surface proteome analysis could be achieved. in combination with label-free quantitative ms, the new protocol can be applied to the differential analysis of the cell surface proteomes between different cell lines to find genetically- or drug-induced changes. we expect this protocol would have broad application in cell surface protein studies, including the discovery of diagnostic marker proteins and potential therapeutic targets.
WOS关键词PLASMA-MEMBRANE PROTEOMICS ; PROTEINS ; IDENTIFICATION ; MCF7 ; PURIFICATION ; NOMENCLATURE ; PEPTIDES ; REVEALS ; MARKER ; LINE
WOS研究方向Biochemistry & Molecular Biology
WOS类目Biochemical Research Methods
语种英语
出版者ELSEVIER SCIENCE BV
WOS记录号WOS:000460716800004
URI标识http://www.irgrid.ac.cn/handle/1471x/2372671
专题大连化学物理研究所
通讯作者Ye, Mingliang
作者单位1.Chinese Acad Sci, Dalian Inst Chem Phys, CAS Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
推荐引用方式
GB/T 7714
Li, Yanan,Wan, Yan,Mao, Jiawei,et al. Sensitive profiling of cell surface proteome by using an optimized biotinylation method[J]. Journal of proteomics,2019,196:33-41.
APA Li, Yanan.,Wan, Yan.,Mao, Jiawei.,Yao, Yating.,Wang, Keyun.,...&Ye, Mingliang.(2019).Sensitive profiling of cell surface proteome by using an optimized biotinylation method.Journal of proteomics,196,33-41.
MLA Li, Yanan,et al."Sensitive profiling of cell surface proteome by using an optimized biotinylation method".Journal of proteomics 196(2019):33-41.

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来源:大连化学物理研究所

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