Proteomics

Dataset Information

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Glyco-CPLL: an integrated method for in-depth and comprehensive N-glycoproteome profiling of human plasma


ABSTRACT: N-glycoproteins are involved in various biological processes. Certain distinctive glycoforms on specific glycoproteins enhance the specificity and/or sensitivity of cancer diagnosis. Therefore, the characterization of plasma N-glycoproteome is essential for new biomarker discovery. Absence of suitable analytical methods for in-depth and large-scale analyses of low-abundance plasma glycoproteins make it challenging to investigate the role of glycosylation. In this study, we developed an integrated method termed Glyco-CPLL, which integrates combinatorial peptide ligand libraries, high-pH reversed-phase pre-fractionation, hydrophilic interaction chromatography, trypsin and PNGase F digestion, shotgun proteomics, and various analysis software (MaxQuant and pGlyco2.0) for the low-abundance plasma glycoproteomic profiling. Then, we utilized the method to perform a comparative study and to explore papillary thyroid carcinoma-related proteins and glycosylations with reference to healthy controls. Finally, a large and comprehensive human plasma N-glycoproteomic database was established, containing 786 proteins, 369 N-glycoproteins, 862 glycosites, 171 glycan compositions, and 1644 unique intact N-glycopeptides. Additionally, several low-abundance plasma glycoproteins were identified, including SVEP1 (~0.54 ng/mL), F8 (~0.83 ng/mL), ADAMTS13 (~1.2 ng/mL). These results suggest that this method will be useful for analyzing plasma intact glycopeptides in future studies. Besides, the Glyco-CPLL method has a great potential to be translated to clinical applications.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood Plasma

SUBMITTER: Yong Zhang  

LAB HEAD: Yong Zhang

PROVIDER: PXD016428 | Pride | 2020-01-09

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
HC-serum-Deglycopeptides-O18-R1.raw Raw
HC-serum-Deglycopeptides-O18-R2.raw Raw
HC-serum-Deglycopeptides-O18-R3.raw Raw
HC-serum-Deglycopeptides-R1.raw Raw
HC-serum-Deglycopeptides-R2.raw Raw
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Publications

Glyco-CPLL: An Integrated Method for In-Depth and Comprehensive N-Glycoproteome Profiling of Human Plasma.

Zhang Yong Y   Mao Yonghong Y   Zhao Wanjun W   Su Tao T   Zhong Yi Y   Fu Linru L   Zhu Jingqiang J   Cheng Jingqiu J   Yang Hao H  

Journal of proteome research 20200114 2


N-glycoproteins are involved in various biological processes. Certain distinctive glycoforms on specific glycoproteins enhance the specificity and/or sensitivity of cancer diagnosis. Therefore, the characterization of plasma N<i>-</i>glycoproteome is essential for a new biomarker discovery. The absence of suitable analytical methods for in-depth and large-scale analyses of low-abundance plasma glycoproteins makes it challenging to investigate the role of glycosylation. In this study, we develope  ...[more]

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