Issue 12, 2022

Chemoproteomic mapping of human milk oligosaccharide (HMO) interactions in cells


Human milk oligosaccharides (HMOs) are a family of unconjugated soluble glycans found in human breast milk that exhibit a myriad of biological activity. While recent studies have uncovered numerous biological functions for HMOs (antimicrobial, anti-inflammatory & probiotic properties), the receptors and protein binding partners involved in these processes are not well characterized. This can be attributed largely in part to the low affinity and transient nature of soluble glycan–protein interactions, precluding the use of traditional characterization techniques to survey binding partners in live cells. Here, we present the use of synthetic photoactivatable HMO probes to capture, enrich and identify HMO protein targets in live cells using mass spectrometry-based chemoproteomics. Following initial validation studies using purified lectins, we profiled the targets of HMO probes in live mouse macrophages. Using this strategy, we mapped hundreds of HMO binding partners across multiple cellular compartments, including many known glycan-binding proteins as well as numerous proteins previously not known to bind glycans. We expect our findings to inform future investigations of the diverse roles of how HMOs may regulate protein function.

Graphical abstract: Chemoproteomic mapping of human milk oligosaccharide (HMO) interactions in cells

Supplementary files

Article information

Article type
01 Aug 2022
09 Oct 2022
First published
18 Oct 2022
This article is Open Access
Creative Commons BY license

RSC Chem. Biol., 2022,3, 1369-1374

Chemoproteomic mapping of human milk oligosaccharide (HMO) interactions in cells

A. A. Hassan, J. M. Wozniak, Z. Vilen, W. Li, A. Jadhav, C. G. Parker and M. L. Huang, RSC Chem. Biol., 2022, 3, 1369 DOI: 10.1039/D2CB00176D

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

Read more about how to correctly acknowledge RSC content.

Social activity