Single Cell Chemical Proteomics with Membrane-Permeable Activity Based Probe for Identification of Functional Proteins in Lysosome of Tumors

被引：17

作者：

Chen, Dongjuan ^{[1
]}

Fan, Fengkai ^{[2
]}

Zhao, Xingfu ^{[1
]}

Xu, Fei ^{[1
]}

Chen, Peng ^{[1
]}

Wang, Jie ^{[1
]}

Ban, Lin ^{[1
]}

Liu, Zhihua ^{[1
]}

Feng, Xiaojun ^{[1
]}

Zhang, Yuhui ^{[1
]}

Liu, Bi-Feng ^{[1
]}

机构：

[1] Huazhong Univ Sci & Technol, Coll Life Sci & Technol,Britton Chance Ctr Biomed, Dept Biomed Engn,Syst Biol Theme, Wuhan Natl Lab Optoelect,Hubei Bioinformat & Mol, Wuhan 430074, Peoples R China

[2] Hubei Univ Educ, Coll Chem & Life Sci, Hubei Key Lab Purificat & Applicat Plant Anticanc, Wuhan 430205, Peoples R China

来源：

ANALYTICAL CHEMISTRY | 2016年 / 88卷 / 04期

基金：

中国国家自然科学基金;

关键词：

INDUCED FLUORESCENCE DETECTION; CAPILLARY-ELECTROPHORESIS; SIGNALING NETWORKS; MASS-SPECTROMETRY; CANCER; CHROMATOGRAPHY; CYTOMETRY; REVEALS; DESIGN;

D O I：

10.1021/acs.analchem.5b04645

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Proteomics at single-cell resolution can help to identify the heterogeneity among cell populations, shows more and more significance in current chemistry and biology. In this work, we demonstrated a new single cell chemical proteomic (SCCP) strategy with a membrane-permeable activity-based probe (ABP) to characterize the functional proteins in lysosome located in the cytosol. The ABP targeted to the cysteine cathepsin family protein, CpFABP-G, was designed for cysteine cathepsins labeling. The labeled HeLa cell of a cancer cell line was injected into a capillary and was lysed by SDS solution with heating. The lysate was then online readout by capillary electrophoresis-laser-induced fluorescence method. Due to the employment of highly specified ABP kicking out the uncorrelated proteins, 4 the expression of cysteine cathepsins in individual HeLa cells was easily 5 detected, and heterogeneity among those HeLa cells was readily discriminated. Further work was concentrated on SCCP analysis of the mouse leukemia cell of monocyte macrophage (RAW264.7). It was for the first time identifying two expression modes of cysteine cathepsins in RAW264.7, which could be undermined by the analysis of cell populations. We believed that SCCP would be one of the powerful alternatives for proteomics at single-cell resolution.

引用

页码：2466 / 2471

页数：6

共 34 条

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