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. 2023 Apr 1;14:1833. doi: 10.1038/s41467-023-37533-6

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a Overview of sialylated (denoted with “S”) epitopes, in the analyzed mucin and released mucin O-glycans. b, c Activity heat maps on sialylated epitopes in released porcine colonic mucin O-glycans (PCM_{free O-glcyans}) and mouse MUC2 attached O-glycans (MUC2_Mouse), respectively after 1 h incubation. d, e Examples showing the broad substrate recognition of AmGH33A and AmGH33B towards α2,3- and α2,6-linked the Neu5Ac and Neu5Gc (light blue) sialic acid forms. f An example illustrating specificity differences between the investigated sialidases. g A comparison of activity profiles on the sum of sialylated extended Tn epitopes either in PCM_{free O-glcyans} and MUC2_Mouse. The enzymatic reactions were performed with PCM_{free O-glcyans} or MUC2_Mouse for 1 h and 24 h. The released PCM O-glycans were compared to non-treated controls, whereas MUC2 O-glycans were released after the enzymatic treatment and compared to non-treated controls. The relative abundances were calculated by integration of the LC-ESI/MS ion chromatogram peak of each glycan and normalising it to the total. The slightly higher relative abundances than the control in some incubations reflect the noise due to small variations in the mucin blots. The relative abundances are the basis for the activity heat map in panel. The “x” marked data are obtained from a single glycan structure, due to the low abundance of the sialyl T epitope in the MUC2_Mouse sample. Sulphatyl substitutions are denoted with “s” in red.