DICP OpenIR
Sensitive profiling of cell surface proteome by using an optimized biotinylation method
Li, Yanan1,2; Wan, Yan1,2; Mao, Jiawei1,2; Yao, Yating1,2; Wang, Keyun1; Qiao, Qinglong1; Fang, Zheng1,2; Ye, Mingliang1
Corresponding AuthorYe, Mingliang(mingliang@dicp.ac.cn)
KeywordCell surface proteins Biotinylation Plasma membrane proteins Proteomics Breast cancer cell lines
Source PublicationJOURNAL OF PROTEOMICS
2019-03-30
ISSN1874-3919
DOI10.1016/j.jprot.2019.01.015
Volume196Pages:33-41
Funding ProjectNational Key R&D Program of China[2017YFA0505003] ; National Key R&D Program of China[2016YFA0501402] ; National Natural Science Foundation of China[21605140] ; National Natural Science Foundation of China[21535008] ; National Natural Science Foundation of China[21405516] ; National Natural Science Foundation of China[21775146] ; National Natural Science Foundation of China[81600046] ; National Natural Science Foundation of China[81430072] ; CAS-Weigao Research & Development Program[[2017]-009] ; DICP, CAS[DICP TMSR201601] ; National Science Fund of China for Distinguished Young Scholars[21525524]
Funding OrganizationNational Key R&D Program of China ; National Key R&D Program of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; CAS-Weigao Research & Development Program ; CAS-Weigao Research & Development Program ; DICP, CAS ; DICP, CAS ; National Science Fund of China for Distinguished Young Scholars ; National Science Fund of China for Distinguished Young Scholars ; National Key R&D Program of China ; National Key R&D Program of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; CAS-Weigao Research & Development Program ; CAS-Weigao Research & Development Program ; DICP, CAS ; DICP, CAS ; National Science Fund of China for Distinguished Young Scholars ; National Science Fund of China for Distinguished Young Scholars ; National Key R&D Program of China ; National Key R&D Program of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; CAS-Weigao Research & Development Program ; CAS-Weigao Research & Development Program ; DICP, CAS ; DICP, CAS ; National Science Fund of China for Distinguished Young Scholars ; National Science Fund of China for Distinguished Young Scholars ; National Key R&D Program of China ; National Key R&D Program of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; CAS-Weigao Research & Development Program ; CAS-Weigao Research & Development Program ; DICP, CAS ; DICP, CAS ; National Science Fund of China for Distinguished Young Scholars ; National Science Fund of China for Distinguished Young Scholars
WOS SubjectBiochemical Research Methods
WOS Research AreaBiochemistry & Molecular Biology
WOS KeywordPLASMA-MEMBRANE PROTEOMICS ; PROTEINS ; IDENTIFICATION ; MCF7 ; PURIFICATION ; NOMENCLATURE ; PEPTIDES ; REVEALS ; MARKER ; LINE
AbstractCell 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.
Language英语
Funding OrganizationNational Key R&D Program of China ; National Key R&D Program of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; CAS-Weigao Research & Development Program ; CAS-Weigao Research & Development Program ; DICP, CAS ; DICP, CAS ; National Science Fund of China for Distinguished Young Scholars ; National Science Fund of China for Distinguished Young Scholars ; National Key R&D Program of China ; National Key R&D Program of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; CAS-Weigao Research & Development Program ; CAS-Weigao Research & Development Program ; DICP, CAS ; DICP, CAS ; National Science Fund of China for Distinguished Young Scholars ; National Science Fund of China for Distinguished Young Scholars ; National Key R&D Program of China ; National Key R&D Program of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; CAS-Weigao Research & Development Program ; CAS-Weigao Research & Development Program ; DICP, CAS ; DICP, CAS ; National Science Fund of China for Distinguished Young Scholars ; National Science Fund of China for Distinguished Young Scholars ; National Key R&D Program of China ; National Key R&D Program of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; CAS-Weigao Research & Development Program ; CAS-Weigao Research & Development Program ; DICP, CAS ; DICP, CAS ; National Science Fund of China for Distinguished Young Scholars ; National Science Fund of China for Distinguished Young Scholars
WOS IDWOS:000460716800004
PublisherELSEVIER SCIENCE BV
Citation statistics
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/165799
Collection中国科学院大连化学物理研究所
Corresponding AuthorYe, Mingliang
Affiliation1.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
Recommended Citation
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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