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A Spin-Tip Enrichment Strategy for the Simultaneous Analysis of N-Glycopeptides and Phosphopeptides from Human Pancreatic Tissues

ABSTRACT: Post-translational modifications (PTMs) change protein structures and functions. Methods for the simultaneous enrichment of multiple PTM types can maximize coverage in analyses. We present a protocol using dual-functional Ti(IV)-immobilized metal affinity chromatography followed by mass spectrometry for the simultaneous enrichment and analysis of protein N-glycosylation and phosphorylation in pancreas tissues.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Pancreas

SUBMITTER: Dylan Nicholas Tabang

LAB HEAD: Lingjun Li

PROVIDER: PXD033065 | Pride | 2022-08-12

REPOSITORIES: Pride

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A Spin-Tip Enrichment Strategy for Simultaneous Analysis of N-Glycopeptides and Phosphopeptides from Human Pancreatic Tissues.

Tabang Dylan Nicholas DN Wang Danqing D Li Lingjun L

Journal of visualized experiments : JoVE 20220504 183

Mass spectrometry can provide deep coverage of post-translational modifications (PTMs), although enrichment of these modifications from complex biological matrices is often necessary due to their low stoichiometry in comparison to non-modified analytes. Most enrichment workflows of PTMs on peptides in bottom-up proteomics workflows, where proteins are enzymatically digested before the resulting peptides are analyzed, only enrich one type of modification. It is the entire complement of PTMs, howe ...[more]

PMID: 35604151

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