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Biological sex is a well-recognised driver of disease susceptibility, drug response, and immune function, yet its influence on protein glycosylation (a post-translational modification critical to protein stability, trafficking, and cell signalling) remains poorly understood across tissues. To address this, Kawahara et al. conducted a comprehensive, multi-omics characterisation of the N-glycoproteome across 19 tissues from adult male and female C57BL/6J mice, integrating quantitative glycoproteomics, proteomics, and transcriptomics with complementary glycomics approaches including lectin microarray profiling, PGC-LC-MS/MS glycomics, and HPLC-based sialic acid speciation.

Peptide and enriched glycopeptide fractions were separated using an IonOpticks Aurora® Ultimate™ 25×75 C18 UHPLC column on a Vanquish Neo interfaced to an Orbitrap Exploris 240. The quantitative glycoproteomics workflow employed TMT-based labelling combined with data-dependent acquisition and stepped HCD fragmentation to identify and quantify over 26,800 unique site-specific N-glycoforms across approximately 1,500 glycoproteins.

Leveraging this workflow, the researchers from the Thaysen-Anderson lab demonstrated that the salivary gland, liver, and kidney display the most pronounced sex-linked differences in glycosylation, driven by coordinated changes in glyco-enzyme expression, particularly sialyltransferases and fucosyltransferases, alongside sex-biased shifts in carrier protein abundance. In contrast, the brain exhibited extensive glycome diversity that was largely conserved between sexes. These findings position glycosylation as a critical, tissue-specific determinant of sex biology, with implications for understanding sex disparities in disease and glycan biomarker discovery.

Publication
bioRxiv

Authors

Rebeca Kawahara, Masaya Hane, Di Wu, Bingyuan Zhang, Fumiya Sakamoto, Takahiro Nakagawa, Takayuki Omoto, Kristina Mae Bienes, Naaz Bansal, Zeynep Sumer-Bayraktar, Sayantani Chatterjee, Koichi Himori, Chiaki Nagai-Okatani, Atsushi Kuno, Makoto Kashima, Daniel Kolarich, Yusuke Matsui, Ken Kitajima, Kenji Kadomatsu, Chihiro Sato, & Morten Thaysen-Andersen;

Title

Multi-omics definition of the sex-specific glycoproteome of murine tissues