An integrated strategy for high-sensitive and multi-level glycoproteome analysis from low micrograms of protein samples

doi:10.1016/j.chroma.2019.04.041

	An integrated strategy for high-sensitive and multi-level glycoproteome analysis from low micrograms of protein samples
	Gao, Weina 3,4,5,6; Li, Hongjie 4,5,6; Liu, Liping 2; Huang, Peiwu 1,4,5; Wang, Zhikun 4,5,6; Chen, Wendong 4,5; Ye, Mingliang 3; Yu, Xiaofang 2; Tian, Ruijun 4,5
刊名	JOURNAL OF CHROMATOGRAPHY A
	2019-08-30
卷号	1600 页码:46-54
关键词	Glycoproteomics Glycopeptide Glycan Integrated sample preparation LC-MS
ISSN号	0021-9673
DOI	10.1016/j.chroma.2019.04.041
通讯作者	Tian, Ruijun(tianrj@sustc.edu.cn)
英文摘要	Glycosylation, as a biologically important protein post-translational modification, often alters on both glycosites and glycans, simultaneously. However, most of current approaches focused on biased profiling of either glycosites or glycans, and limited by time-consuming process and milligrams of starting protein material. We describe here a simple and integrated spintip-based glycoproteomics technology (termed Glyco-SISPROT) for achieving a comprehensive view of glycoproteome with shorter sample processing time and low microgram starting material. By carefully integrating and optimizing SCX, C18 and Concanavalin A (Con A) packing material and their combination in spintip format, both predigested peptides and protein lysates could be processed by Glyco-SISPROT with high efficiency. More importantly, deglycopeptide, intact glycopeptide and glycans released by multiple glycosidases could be readily collected from the same Glyco-SISPROT workflow for LC-MS analysis. In total, above 1850 glycosites in (1) over tilde 770 unique deglycopeptides were characterized from mouse liver by using either 100 mu g of predigested peptides or directly using 100 mu g of protein lysates, in which about 30% of glycosites were released by both PNGase F and Endos. To the best of our knowledge, this approach should be one of the most comprehensive glycoproteomic approaches by using limited protein starting material. One significant benefit of Glyco-SISPROT is that whole processing time is dramatically reduced from a few days to less than 6 h with good reproducibility when protein lysates were directly processed by Glyco-SISPROT. We expect that this method will be suitable for multi-level glycoproteome analysis of rare biological samples with high sensitivity. (C) 2019 Elsevier B.V. All rights reserved.
资助项目	China State Key Basic Research Program Grants[2016YFA0501404] ; China State Key Basic Research Program Grants[2016YFA0501403] ; National Natural Science Foundation of China[21575057] ; Shenzhen Innovation of Science and Technology Commission[JCYJ20160229153100269] ; Shenzhen Innovation of Science and Technology Commission[JCYJ20170412154126026] ; Guangdong Provincial Grants[2016A030312016] ; Guangdong Provincial Grants[2017B030301018]
WOS关键词	N-GLYCOSYLATION SITES ; SELECTIVE ENRICHMENT ; GLYCAN STRUCTURE ; HIGH-MANNOSE ; IDENTIFICATION ; GLYCOPEPTIDES ; QUANTIFICATION ; CHROMATOGRAPHY ; EFFICIENT ; ASSIGNMENT
WOS研究方向	Biochemistry & Molecular Biology ; Chemistry
语种	英语
出版者	ELSEVIER SCIENCE BV
WOS记录号	WOS:000472687800007
资助机构	China State Key Basic Research Program Grants ; China State Key Basic Research Program Grants ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; Shenzhen Innovation of Science and Technology Commission ; Shenzhen Innovation of Science and Technology Commission ; Guangdong Provincial Grants ; Guangdong Provincial Grants ; China State Key Basic Research Program Grants ; China State Key Basic Research Program Grants ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; Shenzhen Innovation of Science and Technology Commission ; Shenzhen Innovation of Science and Technology Commission ; Guangdong Provincial Grants ; Guangdong Provincial Grants ; China State Key Basic Research Program Grants ; China State Key Basic Research Program Grants ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; Shenzhen Innovation of Science and Technology Commission ; Shenzhen Innovation of Science and Technology Commission ; Guangdong Provincial Grants ; Guangdong Provincial Grants ; China State Key Basic Research Program Grants ; China State Key Basic Research Program Grants ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; Shenzhen Innovation of Science and Technology Commission ; Shenzhen Innovation of Science and Technology Commission ; Guangdong Provincial Grants ; Guangdong Provincial Grants
内容类型	期刊论文
源URL	[http://cas-ir.dicp.ac.cn/handle/321008/176173]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
通讯作者	Tian, Ruijun
作者单位	1.Hong Kong Baptist Univ, Dept Chem, State Key Lab Environm & Biol Anal, Hong Kong, Peoples R China 2.Southern Univ Sci & Technol, Affiliated Hosp 1, Shenzhen Peoples Hosp, Shenzhen 518020, Peoples R China 3.Chinese Acad Sci, Dalian Inst Chem Phys, CAS Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China 4.Southern Univ Sci & Technol, Guangdong Prov Key Lab Cell Microenvironm & Dis R, Shenzhen 518055, Peoples R China 5.Southern Univ Sci & Technol, Dept Chem, Shenzhen 518055, Peoples R China 6.Harbin Inst Technol, Sch Chem & Chem Engn, Harbin 150080, Heilongjiang, Peoples R China
推荐引用方式 GB/T 7714	Gao, Weina,Li, Hongjie,Liu, Liping,et al. An integrated strategy for high-sensitive and multi-level glycoproteome analysis from low micrograms of protein samples[J]. JOURNAL OF CHROMATOGRAPHY A,2019,1600:46-54.
APA	Gao, Weina.,Li, Hongjie.,Liu, Liping.,Huang, Peiwu.,Wang, Zhikun.,...&Tian, Ruijun.(2019).An integrated strategy for high-sensitive and multi-level glycoproteome analysis from low micrograms of protein samples.JOURNAL OF CHROMATOGRAPHY A,1600,46-54.
MLA	Gao, Weina,et al."An integrated strategy for high-sensitive and multi-level glycoproteome analysis from low micrograms of protein samples".JOURNAL OF CHROMATOGRAPHY A 1600(2019):46-54.