1. High-throughput N-glycoproteomics with fast liquid chromatographic separation.
- Author
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Bi, Ming and Tian, Zhixin
- Subjects
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LIQUID chromatography , *PROTEIN structure , *PEPTIDES , *HEPATOCELLULAR carcinoma , *LIQUIDS , *MONOSACCHARIDES - Abstract
N -glycosylation is a common protein post translation modification, which has tremendous structure diversity and wide yet delicate regulation of protein structures and functions. Mass spectrometry-based N -glycoproteomics has become a state-of-the-art pipeline for both qualitative and quantitative characterization of N -glycosylation at the intact N -glycopeptide level, providing comprehensive information of peptide backbones, N -glycosites, monosaccharide compositions, sequence and linkage structures. For high-throughput analysis of large-cohort clinic samples, fast and high-performance separation is indispensable. Here we report our development of 1-h liquid chromatography gradient N -glycoproteomics method and accordingly optimized MS parameters. In the benchmark analysis of cancer and paracancerous tissue of hepatocellular carcinoma, 5,218 intact N -glycopeptides were identified, where 422 site- and structure-specific differential N -glycosylation on 145 N -glycoproteins was observed. The method, representing substantial increase of throughput, can be adopted for fast and efficient analysis of N -glycoproteomes at large scale. [Display omitted] • One-hour N -glycoproteomics with 1-h gradient is developed for fast analysis. • 85 % more IDs was achieved per elution time than previous 4-h gradient. • Fast N -glycoproteomics may find wide application of large-cohort study. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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