Evaluation and optimization of high-field asymmetric waveform ion-mobility spectrometry for multiplexed quantitative site-specific N-glycoproteomics
- The heterogeneity and complexity of glycosylation hinder the depth of site-specific glycoproteomics analysis. High-field asymmetric-waveform ion-mobility spectrometry (FAIMS) has been shown to improve the scope of bottom-up proteomics. The benefits of FAIMS for quantitative N-glycoproteomics have not been investigated yet. In this work, we optimized FAIMS settings for N-glycopeptide identification, with or without the tandem mass tag (TMT) label. The optimized FAIMS approach significantly increased the identification of site-specific N-glycopeptides derived from the purified immunoglobulin M (IgM) protein or human lymphoma cells. We explored in detail the changes in FAIMS mobility caused by N-glycopeptides with different characteristics, including TMT labeling, charge state, glycan type, peptide sequence, glycan size, and precursor m/z. Importantly, FAIMS also improved multiplexed N-glycopeptide quantification, both with the standard MS2 acquisition method and with our recently developed Glyco-SPS-MS3 method. The combination of FAIMS and Glyco-SPS-MS3 methods provided the highest quantitative accuracy and precision. Our results demonstrate the advantages of FAIMS for improved mass spectrometry-based qualitative and quantitative N-glycoproteomics.
Author: | Pan Fang, Yanlong JiORCiD, Ivan SilbernORCiDGND, Rosa VinerORCiD, Thomas OellerichORCiD, Kuan-Ting PanORCiD, Henning UrlaubORCiD |
---|---|
URN: | urn:nbn:de:hebis:30:3-629589 |
DOI: | https://doi.org/10.1021/acs.analchem.1c00802 |
ISSN: | 1520-6882 |
Parent Title (English): | Analytical chemistry |
Publisher: | American Chemical Society |
Place of publication: | Columbus, Ohio |
Document Type: | Article |
Language: | English |
Date of Publication (online): | 2021/06/16 |
Date of first Publication: | 2021/06/16 |
Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
Release Date: | 2023/01/27 |
Tag: | Carbohydrates; Chemical biology; Mass spectrometry; Mathematical methods; Peptides and proteins |
Volume: | 93 |
Issue: | 25 |
Page Number: | 10 |
First Page: | 8846 |
Last Page: | 8855 |
HeBIS-PPN: | 507020804 |
Institutes: | Medizin / Medizin |
Dewey Decimal Classification: | 5 Naturwissenschaften und Mathematik / 54 Chemie / 540 Chemie und zugeordnete Wissenschaften |
5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie | |
6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit | |
Sammlungen: | Universitätspublikationen |
Licence (German): | Creative Commons - Namensnennung-Nicht kommerziell - Keine Bearbeitung 4.0 |