A streamlined pipeline for multiplexed quantitative site-specific N-glycoproteomics

2020 | journal article; research paper. A publication with affiliation to the University of Göttingen.

Jump to:Cite & Linked | Documents & Media | Details | Version history

Cite this publication

​A streamlined pipeline for multiplexed quantitative site-specific N-glycoproteomics​
Fang, P.; Ji, Y.; Silbern, I.; Doebele, C. ; Ninov, M.; Lenz, C.   & Oellerich, T. et al.​ (2020) 
Nature Communications11(1).​ DOI: https://doi.org/10.1038/s41467-020-19052-w 

Documents & Media

s41467-020-19052-w.pdf1.29 MBAdobe PDF41467_2020_19052_MOESM1_ESM.pdf2.79 MBAdobe PDF41467_2020_19052_MOESM2_ESM.pdf693.18 kBAdobe PDF41467_2020_19052_MOESM3_ESM.docx13.05 kBMicrosoft Word XML41467_2020_19052_MOESM5_ESM.xlsx778.67 kBMicrosoft Excel XML41467_2020_19052_MOESM6_ESM.xlsx145.23 kBMicrosoft Excel XML41467_2020_19052_MOESM4_ESM.xlsx10.57 MBMicrosoft Excel XML41467_2020_19052_MOESM7_ESM.xlsx98.69 kBMicrosoft Excel XML41467_2020_19052_MOESM8_ESM.xlsx48.87 kBMicrosoft Excel XML41467_2020_19052_MOESM9_ESM.xlsx904.16 kBMicrosoft Excel XML41467_2020_19052_MOESM10_ESM.xlsx13.13 kBMicrosoft Excel XML41467_2020_19052_MOESM11_ESM.pdf147.23 kBAdobe PDF41467_2020_19052_MOESM12_ESM.xlsx7.69 MBMicrosoft Excel XML

License

Published Version

Attribution 4.0 CC BY 4.0

Details

Authors
Fang, Pan; Ji, Yanlong; Silbern, Ivan; Doebele, Carmen ; Ninov, Momchil; Lenz, Christof ; Oellerich, Thomas; Pan, Kuan-Ting; Urlaub, Henning 
Abstract
Regulation of protein N-glycosylation is essential in human cells. However, large-scale, accurate, and site-specific quantification of glycosylation is still technically challenging. We here introduce SugarQuant, an integrated mass spectrometry-based pipeline comprising protein aggregation capture (PAC)-based sample preparation, multi-notch MS3 acquisition (Glyco-SPS-MS3) and a data-processing tool (GlycoBinder) that enables confident identification and quantification of intact glycopeptides in complex biological samples. PAC significantly reduces sample-handling time without compromising sensitivity. Glyco-SPS-MS3 combines high-resolution MS2 and MS3 scans, resulting in enhanced reporter signals of isobaric mass tags, improved detection of N-glycopeptide fragments, and lowered interference in multiplexed quantification. GlycoBinder enables streamlined processing of Glyco-SPS-MS3 data, followed by a two-step database search, which increases the identification rates of glycopeptides by 22% compared with conventional strategies. We apply SugarQuant to identify and quantify more than 5,000 unique glycoforms in Burkitt’s lymphoma cells, and determine site-specific glycosylation changes that occurred upon inhibition of fucosylation at high confidence.
Issue Date
2020
Journal
Nature Communications 
Project
SFB 1286: Quantitative Synaptologie 
SFB 1286 | A06: Mitochondrienfunktion und -umsatz in Synapsen 
Working Group
RG Urlaub (Bioanalytische Massenspektrometrie) 
External URL
https://sfb1286.uni-goettingen.de/literature/publications/59
eISSN
2041-1723
Language
English

Reference

Citations


Social Media