A Fully Integrated Spintip-Based Approach for Sensitive and Quantitative Profiling of Region-Resolved in Vivo Brain Glycoproteome

Huang, Peiwu1,2; Li, Hongjie1; Gao, Weina1; Cai, Zongwei2; Tian, Ruijun1

2019-07-16

10.1021/acs.analchem.9b01930

ANALYTICAL CHEMISTRY   影响因子和分区

0003-2700

1520-6882

Region- and cell type-resolved global proteome and specific post-translational modifications (PTMs) profiling of tissues has drawn great attention recently for interpreting the heterogeneous multicellular microenvironment of various in vivo systems. Due to access to low microgram of proteins and low abundance of glycoproteins, spatially resolved glycoproteome analysis of in vivo tissue sections remains challenging. Several glycoproteomics sample preparation strategies were established for processing microgram-level of protein samples, but these strategies were not either fully integrated or directly compatible with tissue samples when considering protein extraction in strong lysis buffers. Moreover, these approaches mainly focused on identification of glycosylation sites, but pay less attention to quantification, all of which limit their applications. Here we designed a fully integrated spintip-based glycoproteomic approach (FISGlyco) which achieves all the steps of glycoprotein enrichment, digestion, deglycosylation, and desalting in single spintip device. Sample loss is significantly reduced, and the total processing time is reduced to 4 h, while detection sensitivity and label-free quantification precision is greatly improved. 607 N-glycosylation sites were successfully identified and quantified from only 5 mu g of mouse brain proteins. By seamlessly combining with laser capture microdissection (LCM), the first region-resolved N-glycoproteome profiling of four mouse brain regions, including isocortex, hippocampus, thalamus, and hypothalamus, was achieved, with 1875, 1794, 1801, and 1417 N-glycosites identified, respectively. Our approach could be a generic approach for region and even cell type specific glycoproteome analysis of in vivo tissue sections.

SCI ; EI

英语

NI期刊

第一 ; 通讯

Guangdong Provincial Grants[2016A030312016] ; Guangdong Provincial Grants[2017B030301018]

Chemistry

Chemistry, Analytical

WOS:000476567700061

AMER CHEMICAL SOC

20193207292740

Brain ; Desalination ; Glycoproteins ; Glycosylation ; Microdissection ; Proteins ; Tissue

Water Treatment Techniques:445.1 ; Bioengineering and Biology:461 ; Organic Compounds:804.1

CHEMISTRY

Web of Science

1.Southern Univ Sci & Technol, Dept Chem, Shenzhen 518055, Peoples R China
2.Hong Kong Baptist Univ, Dept Chem, State Key Lab Environm & Biol Anal, Hong Kong, Peoples R China

Huang, Peiwu,Li, Hongjie,Gao, Weina,et al. A Fully Integrated Spintip-Based Approach for Sensitive and Quantitative Profiling of Region-Resolved in Vivo Brain Glycoproteome[J]. ANALYTICAL CHEMISTRY,2019,91(14):9181-9189.

Huang, Peiwu,Li, Hongjie,Gao, Weina,Cai, Zongwei,&Tian, Ruijun.(2019).A Fully Integrated Spintip-Based Approach for Sensitive and Quantitative Profiling of Region-Resolved in Vivo Brain Glycoproteome.ANALYTICAL CHEMISTRY,91(14),9181-9189.

Huang, Peiwu,et al."A Fully Integrated Spintip-Based Approach for Sensitive and Quantitative Profiling of Region-Resolved in Vivo Brain Glycoproteome".ANALYTICAL CHEMISTRY 91.14(2019):9181-9189.