N-glycans of bovine submaxillary mucin contain core-fucosylated and sulfated glycans but not sialylated glycans

Kim, Jihye; Lee, Junmyoung; Jang, Yeonjoo; Ha, Jongkwan; Kim, Donghwi; Ji, Minkyoo; Lee, Young Kwang; Kim, Woo Seok; You, Seungkwan; Do, Jonghye; Ryu, Changsoo; Kim, Ha Hyung

doi:10.1016/j.ijbiomac.2019.07.108

Cited by 14 publications

(6 citation statements)

References 39 publications

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“…The bovine submaxillary mucins (BSMs) used herein contain about 50% sialyl Tn and 10% Tn antigens. The sialic acid residues, composed of ∼70% Neu5Ac and ∼30% Neu5Gc can make up to 30% of the molecule’s mass. , A number of non-sialylated N -glycans are also present on the BSM . Sialic acids are of particular interest in this study since they play important roles in immunity.…”

Section: Discussionmentioning

confidence: 99%

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Glyco-Modification of Mucin Hydrogels to Investigate Their Immune Activity

Yan

Hjorth

Winkeljann

et al. 2020

ACS Appl. Mater. Interfaces

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Mucins are multifunctional glycosylated proteins that are increasingly investigated as building blocks of novel biomaterials. An attractive feature is their ability to modulate the immune response, in part by engaging with sialic acid binding receptors on immune cells. Once assembled into hydrogels, bovine submaxillary mucins (Muc gels) were shown to modulate the recruitment and activation of immune cells and avoid fibrous encapsulation in vivo. However, nothing is known about the early immune response to Muc gels. This study characterizes the response of macrophages, important orchestrators of the material-mediated immune response, over the first 7 days in contact with Muc gels. The role of mucin-bound sialic acid sugar residues was investigated by first enzymatically cleaving the sugar and then assembling the mucin variants into covalently cross-linked hydrogels with rheological and surface nanomechanical properties similar to nonmodified Muc gels. Results with THP-1 and human primary peripheral blood monocytes derived macrophages showed that Muc gels transiently activate the expression of both pro-inflammatory and anti-inflammatory cytokines and cell surface markers, for most makers with a maximum on the first day and loss of the effect after 7 days. The activation was sialic acid-dependent for a majority of the markers followed. The pattern of gene expression, protein expression, and functional measurements did not strictly correspond to M1 or M2 macrophage phenotypes. This study highlights the complex early events in macrophage activation in contact with mucin materials and the importance of sialic acid residues in such a response. The enzymatic glyco-modulation of Muc gels appears as a useful tool to help understand the biological functions of specific glycans on mucins which can further inform on their use in various biomedical applications.

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Section: Discussionmentioning

confidence: 99%

“…40,41 A number of nonsialylated N-glycans are also present on the BSM. 42 Sialic acids are of particular interest in this study since they play important roles in immunity. Physically, owing to their localization at the tip of the glycosylation and their negative charge, sialic acid mediate cell−cell interactions and mask antigens.…”

Section: ■ Discussionmentioning

confidence: 99%

Glyco-Modification of Mucin Hydrogels to Investigate Their Immune Activity

Yan

Hjorth

Winkeljann

et al. 2020

ACS Appl. Mater. Interfaces

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“…Schematic illustrations of the mucin structures based on the present results and previous reports [12,14] are shown in Figure 4. The N-glycans, including the negatively charged sulfated N-glycans identified in this study, are located in the hydrophobic terminal regions ( Figure 4A) [12,19]. The negatively charged glycans or positively charged amino acids (containing Lys, Arg, and His) in mucin might enhance its adsorption onto positive or negative surfaces in an aqueous environment, respectively.…”

Section: Discussionmentioning

confidence: 80%

“…BSM protein purity was confirmed using SDS-PAGE (data not shown), as described in our previous reports [19,29]. The N-glycans released from BSM were derivatized with ProA and analyzed using UPLC and LC-ESI-HCD-MS/MS.…”

Section: Uplc and Lc-esi-hcd-ms/ms Analysis Of N-glycansmentioning

confidence: 91%

N-Glycan Modifications with Negative Charge in a Natural Polymer Mucin from Bovine Submaxillary Glands, and Their Structural Role

Kim

Lee

et al. 2020

Polymers

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Bovine submaxillary mucin (BSM) is a natural polymer used in biomaterial applications for its viscoelasticity, lubricity, biocompatibility, and biodegradability. N-glycans are important for mucin stability and function, but their structures have not been fully characterized, unlike that of O-glycans. In this study, BSM N-glycans were investigated using liquid chromatography-tandem mass spectrometry. The microheterogeneous structures of 32 N-glycans were identified, and the quantities (%) of each N-glycan relative to total N-glycans (100%) were obtained. The terminal N-acetylgalactosamines in 12 N-glycans (sum of relative quantities; 27.9%) were modified with mono- (10 glycans) and disulfations (2 glycans). Total concentration of all sulfated N-glycans was 6.1 pmol in BSM (20 µg), corresponding to 25.3% of all negatively charged glycans (sum of present N-glycans and reported O-glycans). No N-glycans with sialylated or phosphorylated forms were identified, and sulfate modification ions were the only negative charges in BSM N-glycans. Mucin structures, including sulfated N-glycans located in the hydrophobic terminal regions, were indicated. This is the first study to identify the structures and quantities of 12 sulfated N-glycans in natural mucins. These sulfations play important structural roles in hydration, viscoelasticity control, protection from bacterial sialidases, and polymer stabilization to support the functionality of BSM via electrostatic interactions.

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“…Sulfated N-glycans have been observed in a wide variety of organisms ranging from animals to viruses (Fig. S13) (23)(24)(25)(26). These decorations can take the form of GalNAc-6S, GalNAc-4S, Gal-3S, Gal-6S, and Man-6S (36).…”

Section: B and Fig S6)mentioning

confidence: 99%

Plant N-glycan breakdown by human gut Bacteroides

Crouch

Urbanowicz

Baslé

et al. 2022

Preprint

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The major nutrients available to the human colonic microbiota are complex glycans derived from the diet. To degrade this highly variable mix of sugar structures, gut microbes have acquired a huge array of different carbohydrate-active enzymes (CAZymes), predominantly glycoside hydrolases, many of which have specificities that can be exploited for a range of different applications. Plant N-glycans are prevalent on proteins produced by plants and thus components of the diet, but the breakdown of these complex molecules by the gut microbiota has not been explored. Plant N-glycans are also well characterised allergens in pollen and some plant-based foods, and when plants are used in heterologous protein production for medical applications, the N-glycans present can pose a risk to therapeutic function and stability. Here we use a novel genome association approach for enzyme discovery to identify a breakdown pathway for plant complex N-glycans encoded by a gut Bacteroides species and biochemically characterise five CAZymes involved, including structures of the PNGase and GH92 α-mannosidase. These enzymes provide a toolbox for the modification of plant N-glycans for a range of potential applications. Furthermore, the keystone PNGase also has activity against insect-type N-glycans, which we discuss from the perspective of insects as a nutrient source.

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N-glycans of bovine submaxillary mucin contain core-fucosylated and sulfated glycans but not sialylated glycans

Cited by 14 publications

References 39 publications

Glyco-Modification of Mucin Hydrogels to Investigate Their Immune Activity

Glyco-Modification of Mucin Hydrogels to Investigate Their Immune Activity

N-Glycan Modifications with Negative Charge in a Natural Polymer Mucin from Bovine Submaxillary Glands, and Their Structural Role

Plant N-glycan breakdown by human gut Bacteroides

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