ABSTRACT
Ubiquitous occurrence in Nature, abundant presence at strategically important places such as the cell surface and dynamic shifts in their profile by diverse molecular switches qualifies the glycans to serve as versatile biochemical signals. However, their exceptional structural complexity often prevents one noting how simple the rules of objective-driven assembly of glycan-encoded messages are. This review is intended to provide a tutorial for a broad readership. The principles of why carbohydrates meet all demands to be the coding section of an information transfer system, and this at unsurpassed high density, are explained. Despite appearing to be a random assortment of sugars and their substitutions, seemingly subtle structural variations in glycan chains by a sophisticated enzymatic machinery have emerged to account for their specific biological meaning. Acting as 'readers' of glycan-encoded information, carbohydrate-specific receptors (lectins) are a means to turn the glycans' potential to serve as signals into a multitude of (patho)physiologically relevant responses. Once the far-reaching significance of this type of functional pairing has become clear, the various modes of spatial presentation of glycans and of carbohydrate recognition domains in lectins can be explored and rationalized. These discoveries are continuously revealing the intricacies of mutually adaptable routes to achieve essential selectivity and specificity. Equipped with these insights, readers will gain a fundamental understanding why carbohydrates form the third alphabet of life, joining the ranks of nucleotides and amino acids, and will also become aware of the importance of cellular communication via glycan-lectin recognition.
Subject(s)
Carbohydrate Metabolism , Carbohydrates , Lectins , Signal Transduction/physiology , Animals , Carbohydrates/chemistry , Carbohydrates/genetics , Humans , Lectins/chemistry , Lectins/genetics , Lectins/metabolismABSTRACT
In this review, we document the evolution of common glycan structures in the eukaryotes, and illustrate the considerable variety of oligosaccharides existing in these organisms. We focus on the families of N- and O-glycans, glycosphingolipids, glycosaminoglycans, glycosylphosphatidylinositol (GPI) anchors, sialic acids (Sias), and cytoplasmic and nuclear glycans. We also outline similar and divergent aspects of the glycans during evolution within the groups, which include inter- and intraspecies differences, molecular mimicry, viral glycosylation adaptations, glycosyltransferase specificity relating to function, and the natural dynamism powering these events. Finally, we present an overview of the patterns of glycosylation found within the groups comprising the Eukaryota, namely the Deuterostomia, Fungi, Viridiplantae, Nematoda, and Arthropoda.
Subject(s)
Polysaccharides/physiology , Protein Processing, Post-Translational , Animals , Carbohydrate Conformation , Carbohydrate Sequence , Evolution, Molecular , Glycoproteins/metabolism , Glycosylation , Humans , Molecular Mimicry , Molecular Sequence DataABSTRACT
BACKGROUND: The mucins found as components of mucus gel layers at mucosal surfaces throughout the body play roles in protection as part of the defensive barrier on an organ and tissue specific basis. SCOPE OF THE REVIEW: The human MUC gene family codes up to 20 known proteins, which can be divided into secreted and membrane-associated forms each with a typical protein domain structure. The secreted mucins are adapted to cross link in order to allow formation of the extended mucin networks found in the secreted mucus gels. The membrane-associated mucins possess membrane specific domains which enable their various biological functions as part of the glycocalyx. All mucins are highly O-glycosylated and this is tissue specific and linked with specific biological functions at these locations. Mucin biology is dynamic and the processes of degradation and turnover are well integrated with biosynthesis to maintain a continuous mucosal protection against all external aggressive forces. Interaction of mucins with microflora plays an important role in normal function. Mucins are modified in a variety of diseases and this may be due to abberant mucin peptide or glycosylation. MAJOR CONCLUSIONS: Mucins represent a family of glycoprotein having fundamental roles in mucosal protection and communication with external environment. GENERAL SIGNIFICANCE: The review emphasises the nature of mucins as glycoproteins and their role in presenting an array of glycan structures at the mucosal cell surface.
Subject(s)
Carbohydrates/chemistry , Mucins/chemistry , Mucins/metabolism , Mucous Membrane/metabolism , Enterobacteriaceae/metabolism , Enterobacteriaceae/physiology , Host-Pathogen Interactions , Humans , Models, Biological , Mucins/genetics , Mucous Membrane/microbiology , Multigene Family/genetics , Neoplasms/metabolism , Polysaccharides/chemistryABSTRACT
The natural glycopeptide antibiotic teicoplanin is used for the treatment of serious Gram-positive related bacterial infections and can be administered intravenously, intramuscularly, topically (ocular infections), or orally. It has also been considered for targeting viral infection by SARS-CoV-2. The hydrodynamic properties of teicoplanin A2 (M1 = 1880 g/mol) were examined in phosphate chloride buffer (pH 6.8, I = 0.10 M) using sedimentation velocity and sedimentation equilibrium in the analytical ultracentrifuge together with capillary (rolling ball) viscometry. In the concentration range, 0-10 mg/mL teicoplanin A2 was found to self-associate plateauing > 1 mg/mL to give a molar mass of (35,400 ± 1000) g/mol corresponding to ~ (19 ± 1) mers, with a sedimentation coefficient s20, w = ~ 4.65 S. The intrinsic viscosity [[Formula: see text]] was found to be (3.2 ± 0.1) mL/g: both this, the value for s20,w and the hydrodynamic radius from dynamic light scattering are consistent with a globular macromolecular assembly, with a swelling ratio through dynamic hydration processes of ~ 2.
Subject(s)
COVID-19 , Teicoplanin , Humans , Hydrodynamics , SARS-CoV-2 , Anti-Bacterial Agents , GlycopeptidesABSTRACT
Glycopeptide antibiotics are regularly used in ophthalmology to treat infections of Gram-positive bacteria. Aggregative interactions of antibiotics with mucins however can lead to long exposure and increases the risk of resistant species. This study focuses on the evaluation of potential interactions of the last line of defence glycopeptide antibiotic teicoplanin with an ocular mucin model using precision matrix free hydrodynamic and microscopic techniques: sedimentation velocity in the analytical ultracentrifuge (SV-AUC), dynamic light scattering (DLS) and atomic force microscopy (AFM). For the mixtures of teicoplanin at higher doses (1.25 mg/mL and 12.5 mg/mL), it was shown to interact and aggregate with bovine submaxillary mucin (BSM) in the distributions of both sedimentation coefficients by SV-AUC and hydrodynamic radii by DLS. The presence of aggregates was confirmed by AFM for higher concentrations. We suggest that teicoplanin eye drop formulations should be delivered at concentrations of < 1.25 mg/mL to avoid potentially harmful aggregations.
Subject(s)
Hydrodynamics , Teicoplanin , Animals , Cattle , Mucins , Anti-Bacterial Agents/pharmacology , GlycopeptidesABSTRACT
IBDs (inflammatory bowel diseases) are a group of diseases affecting the gastrointestinal tract. The diseases are multifactorial and cover genetic aspects: susceptibility genes, innate and adaptive responses to inflammation, and structure and efficacy of the mucosal protective barrier. Animal models of IBD have been developed to gain further knowledge of the disease mechanisms. These topics form an overlapping background to enable an improved understanding of the molecular features of these diseases. A series of articles is presented based on the topics covered at the Biochemical Society Focused Meeting The Molecular Biology of Inflammatory Bowel Diseases.
Subject(s)
Inflammatory Bowel Diseases/genetics , Animals , Diet , Gastrointestinal Tract/microbiology , Gastrointestinal Tract/pathology , Gastrointestinal Tract/physiopathology , Humans , Inflammatory Bowel Diseases/etiology , Inflammatory Bowel Diseases/microbiology , Inflammatory Bowel Diseases/physiopathology , Molecular BiologyABSTRACT
BACKGROUND: Mucositis is a painful ulcerative condition of the oral cavity and gastrointestinal tract, occurring in association with chemotherapy and radiotherapy regimes. Trefoil factor family peptides (TFF, trefoil peptides), present in saliva, contribute to epithelial restitution and repair and are therefore potentially important in the healing phase of mucositis. This study aimed to assess any changes in the levels of trefoil peptides in oncology patients with and without mucositis. METHODS: Saliva was collected from healthy children, pretreatment oncology patients, neutropenic patients on treatment with no oral disease and mucositic patients. TFF1, 2 and 3 were quantified using ELISA. RESULTS: In healthy children TFF2 and 3 were positively correlated with age (r = 0.454, p = 0.01 for TFF2; r = 0.410, p = 0.05 for TFF3 Spearman rank correlation). TFF3 was higher in mucositis compared to all other groups. A linear regression prediction model indicated that TFF3, but not TFF1 and TFF2, was significantly different in mucositic and healthy controls, suggesting an altered pattern of trefoil peptide secretion (p = 0.021). CONCLUSIONS: This study is the first to focus on trefoil peptides in paediatric saliva. It shows the correlation between TFF2, TFF3 and age in healthy children. Paediatric mucositis disease occurs in the presence of increased concentrations and an altered pattern of trefoil peptides.
Subject(s)
Mucositis/metabolism , Peptides/analysis , Saliva/metabolism , Tumor Suppressor Proteins/analysis , Adolescent , Child , Child, Preschool , Enzyme-Linked Immunosorbent Assay , Female , Humans , Infant , Male , Mucositis/pathology , Trefoil Factor-1 , Trefoil Factor-2 , Trefoil Factor-3ABSTRACT
This is the first differential expression proteomics study on a human syngeneic cellular in vitro progression model of the colorectal adenoma-to-carcinoma sequence, the anchorage-dependent non-tumorigenic adenoma derived cell line AA/C1 and the derived anchorage-independent and tumorigenic carcinoma cell line AA/C1/SB10C. The study is based on quantitative 2-DE and is complemented by Western blot validation. Excluding redundancies due to proteolysis and post-translational modified isoforms of over 2000 protein spots, 13 proteins were revealed as regulated with statistical variance being within the 95th confidence level and were identified by peptide mass fingerprinting in MALDI MS. Progression-associated proteins belong to the functional complexes of anaerobic glycolysis/gluconeogenesis, steroid biosynthesis, prostaglandin biosynthesis, the regulation and maintenance of the cytoskeleton, protein biosynthesis and degradation, the regulation of apoptosis or other functions. Partial but significant overlap was revealed with previous proteomics and transcriptomics studies in colorectal carcinoma. Among upregulated proteins we identified 3-HMG-CoA synthase, protein phosphatase 1, prostaglandin E synthase 2, villin 1, annexin A1, triosephosphate isomerase, phosphoserine aminotransferase 1, fumarylacetoacetate hydrolase and pyrroline-5-carboxylate reductase 1 (PYCR1), while glucose-regulated protein 78, cathepsin D, lamin A/C and quinolate phosphoribosyltransferase were downregulated.
Subject(s)
Adenoma/chemistry , Adenoma/pathology , Colorectal Neoplasms/chemistry , Colorectal Neoplasms/pathology , Disease Progression , Proteome/analysis , Cell Line, Tumor , Down-Regulation , Electrophoresis, Gel, Two-Dimensional , Humans , Models, Biological , Proteomics , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Up-RegulationABSTRACT
The glycans (carbohydrates) form a diverse group of biomolecules which play active parts in most physiological processes. The field of structural glycobiology concerns the structures of the glycans themselves, the proteins which interact with them and the nature of the interactions between the two. The resulting information is important for our understanding of human health and disease, and for development of new therapeutic strategies. A series of articles is introduced based on the topics covered at the Structural Glycobiology and Human Health Biochemical Society Focused Meeting. Their subjects range from in-depth determinations of three-dimensional protein structure to broad screening techniques for glycan-protein interactions relevant to disease processes, including bacterial, parasitic and viral infections, inflammatory processes, cancer and diabetes.
Subject(s)
Glycomics , Acetylglucosamine/metabolism , Glycosaminoglycans/metabolism , Heparitin Sulfate/metabolism , Humans , Lipopolysaccharides/metabolism , Systems BiologyABSTRACT
The present article provides an overview on mucins and their role in biological processes, while aiming to familiarize readers with the current tools available for the synthesis of structurally defined mucin-type glycan probes including the advantages and potential applications of using ionic liquids in the synthesis of this important class of oligosaccharides. Furthermore, we also highlight recent developments in glycoarray technology that can enable high-sensitivity and high-throughput analysis of this important class of protein-carbohydrate interactions.
Subject(s)
Ionic Liquids/chemistry , Mucins/chemistry , Oligosaccharides/chemistry , Oligosaccharides/chemical synthesis , Polysaccharides/chemistry , Models, ChemicalABSTRACT
Glycoproteins are major players in the mucus protective barrier in the gastrointestinal and other mucosal surfaces. In particular the mucus glycoproteins, or mucins, are responsible for the protective gel barrier. They are characterized by their high carbohydrate content, present in their variable number, tandem repeat domains. Throughout evolution the mucins have been maintained as integral components of the mucosal barrier, emphasizing their essential biological status. The glycosylation of the mucins is achieved through a series of biosynthetic pathways processes, which generate the wide range of glycans found in these molecules. Thus mucins are decorated with molecules having information in the form of a glycocode. The enteric microbiota interacts with the mucosal mucus barrier in a variety of ways in order to fulfill its many normal processes. How bacteria read the glycocode and link to normal and pathological processes is outlined in the review.
ABSTRACT
Mucin glycoproteins and trefoil peptides play an important role in protection and repair of the gastrointestinal epithelium. This study investigates alterations in mucin and trefoil peptide gene expression and product localization in ulcerative colitis (UC). Product localization and message expression of mucin MUC1 to 6 and trefoil peptide TFF1 to 3 genes was analyzed in rectosigmoid tissue from a cohort of patients with active UC and compared with that of normal colorectal mucosa. MUC1 expression was upregulated in severe UC at the site of rupture of crypt abscesses. Reduction in MUC2 expression occurred in UC adjacent to ulceration. No alteration in MUC3 or MUC4 gene expression was detectable in UC compared with normal colorectal mucosa. No ectopic expression of MUC5AC, MUC5B, or MUC6 was identified in UC. Ectopic TFF1 expression was identified in tissues eliciting histological features of severe disease. Decreased TFF3 localization was demonstrated in UC tissues, but no TFF2 expression was detected in any colorectal specimens. Subtle alterations in composition of the supramucosal defense barrier exist in UC and vary in relation to clinical severity of disease. There is upregulation in mucin MUC1 at crypt abscesses and neo-expression of TFF1 trefoil peptide in severe disease.
Subject(s)
Colitis, Ulcerative/pathology , Gastric Mucosa/pathology , Glycoproteins/genetics , Mucins/genetics , Peptides/genetics , Adult , Aged , Aged, 80 and over , Cohort Studies , Female , Gene Expression Regulation/immunology , Glycoproteins/metabolism , Humans , Immunity, Mucosal , Immunohistochemistry , Male , Middle Aged , Mucins/metabolism , Peptides/metabolism , Prospective Studies , RNA, Messenger/genetics , Severity of Illness Index , Trefoil Factor-2ABSTRACT
PURPOSE: Contact lens wear alters the preocular fluid through factors that include tear deposits on the lens. In the current study, lens-adherent material was extracted to assess whether contact lenses sample mucins from the preocular fluid. METHODS: Discarded extended-wear contact lenses were collected from patients with no ocular surface disease. Mucins were extracted in guanidine hydrochloride (GuHCl) with protease inhibitors. After the supernatant was removed, the extraction was repeated twice with the addition of 10 mM dithiothreitol, making a total of three extractions. Mucins were isolated by cesium chloride (CsCl) gradient centrifugation and size fractionated on Sepharose CL2B. Charge distribution was analyzed on ion-exchange chromatography with a lithium perchlorate (LiClO(4)) gradient. RESULTS: Contact lens-adherent mucins comprised soluble mucins and mucins that required solubilization by (repeated) dithiothreitol treatment. MUC1, MUC4, MUC2, and MUC5AC mucins eluted mainly at low buoyant densities in extractions from lenses worn long term without disinfection and at successively higher buoyant densities from monthly disposable contact lenses. Mucins with little negative charge, which were observed in all extractions, and very highly negatively charged species, present in the second and third extractions from contact lenses, had no equivalents in tissue-extracted mucins. CONCLUSIONS: Mucins adhering to contact lenses are altered forms of intracellular mucins. Different degrees of adherence of mucins to contact lenses may occur, either because of mucin characteristics or after mucin complexation with adherent materials. In the context of good contact lens hygiene, their presence may offer some protection from toxicants in the tear film, because mucins could function as acceptors for charged moieties such as free radicals.
Subject(s)
Contact Lenses, Extended-Wear , Eye Proteins/metabolism , Mucins/metabolism , Chromatography, Ion Exchange , Disposable Equipment , Eye Proteins/analysis , Eye Proteins/isolation & purification , Female , Humans , Mucins/analysis , Mucins/isolation & purification , Protein Binding , Tears/metabolismABSTRACT
PURPOSE: Mucins are polymers that may reduce drag and enhance tear outflow. Mucin expression and distribution in human efferent tear ducts were tested in the physiological state, and potential differences in the expression pattern were investigated in the presence of primary acquired dacryostenosis (PANDO). METHODS: Expression of mucins in human lacrimal sac and nasolacrimal ducts was monitored by reverse transcription-polymerase chain reaction analysis. The presence and distribution of MUC1, -2, -4, -5AC, -5B, -6, and -7 in epithelia of the efferent tear duct passage are assessed with antisera to mucin peptide cores. Twenty normal tissues from cadavers and surgical specimens from 20 patients with PANDO were tested. RESULTS: mRNAs for all mucins investigated were detected in healthy human lacrimal sacs and nasolacrimal ducts. MUC6 mRNA was detected in only about half of the investigated samples. A reduced level of MUC2, -5AC, and -5B mRNAs was observed in PANDO. Immunohistochemistry revealed MUC2 in goblet cells and single epithelial cells. Both MUC5AC and -5B were detected in goblet cells forming intraepithelial mucous glands. MUC7 was present only in columnar epithelial cells of the efferent tear duct system. No immunoreactivity was observed with antibodies against MUC1, -4, and -6 peptide cores. CONCLUSIONS: Human efferent tear ducts express and produce a broad spectrum of mucins that is partly comparable with that in the conjunctiva and the salivary glands. The mucin diversity of the efferent tear ducts could enhance tear transport and antimicrobial defense. Reduced levels of mucin mRNA in a nonfunctioning though patent segment of the lacrimal passage, which is associated with epiphora, suggests that mucins ease tear flow through the efferent tear ducts.
Subject(s)
Lacrimal Duct Obstruction/metabolism , Mucins/genetics , Mucins/metabolism , Nasolacrimal Duct/metabolism , Adult , Aged , Aged, 80 and over , DNA, Complementary/biosynthesis , Dacryocystorhinostomy , Epithelial Cells/metabolism , Female , Goblet Cells/metabolism , Humans , Immunoenzyme Techniques , Lacrimal Apparatus/metabolism , Male , Middle Aged , RNA/isolation & purification , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
OBJECTIVE: Airway mucins may play an important role in the mechanism of respiratory complications after cardiopulmonary bypass in infants and children. Our aim was to measure airway mucin levels before and after cardiopulmonary bypass and to determine whether changes in mucin levels were associated with the development of respiratory complications. METHODS: Airway glycoprotein and mucins (MUC5AC, MUC5B, and MUC2) in serial small-volume airway lavage samples from 39 young children who underwent cardiac operations with cardiopulmonary bypass were measured by slot-blot assay with specific antimucin peptide antibodies. The relationship between mucin changes and post-cardiopulmonary bypass respiratory complications was investigated. Airway lavage samples were also collected from 11 children before and after operation without cardiopulmonary bypass, and changes in mucin levels were compared with those in subjects who underwent cardiopulmonary bypass. Airway lavage sample DNA was also measured to investigate the relationship between mucin changes and lung injury. RESULTS: Glycoprotein, MUC5AC, and MUC5B levels were significantly increased after cardiopulmonary bypass (P <.001) whereas MUC2 level was not. Children with respiratory complications showed significantly higher glycoprotein and MUC5AC levels than did children without respiratory complications before and after cardiopulmonary bypass (P <.05). Increase of total mucin (MUC5AC, MUC5B, and MUC2) during cardiopulmonary bypass showed positive correlation with DNA increase during cardiopulmonary bypass (r = 0.73), PaCO(2) (r = 0.62) and alveolar-arterial oxygen difference (r = 0.55) immediately after cardiopulmonary bypass. Increase of total mucin was associated with postoperative respiratory complications and their severity. There were no significant changes detected in airway mucin during operations without cardiopulmonary bypass. CONCLUSIONS: Airway mucins were increased during cardiopulmonary bypass, and this increase was associated with markers of lung injury after cardiopulmonary bypass and with the development of postoperative respiratory complications.
Subject(s)
Cardiopulmonary Bypass , Mucins/metabolism , Postoperative Complications/etiology , Postoperative Complications/metabolism , Respiratory Distress Syndrome/etiology , Respiratory Distress Syndrome/metabolism , Trachea/metabolism , Biomarkers/analysis , Bronchoalveolar Lavage Fluid/chemistry , Cardiac Surgical Procedures , Child , Child Welfare , Child, Preschool , Glycoproteins/metabolism , Heart Defects, Congenital/metabolism , Heart Defects, Congenital/surgery , Humans , Infant , Infant Welfare , Mucins/classification , Oxygen/blood , Postoperative Period , Respiratory Mucosa/metabolism , Severity of Illness Index , Statistics as Topic , Time Factors , Treatment Outcome , United KingdomABSTRACT
AIM: The supramucosal gel, crucial for gut barrier function, might be compromised in necrotizing enterocolitis (NEC). Breast milk is associated with a reduced incidence of NEC. We compared the effects of human breast milk (BM) versus a neonatal formula, Nutriprem 1 (FF), on adherence, internalisation, and penetration of NEC-associated Escherichia coli through monolayers of mucus producing intestinal cells, HT29-MTX-E12 (E12). METHODS: E12 cells were grown to confluence on membranes permeable to bacteria. E. coli, reference strain and isolated from a NEC-affected intestine, were cultured in LB broth, labelled with fluorescein and biotinylated. Bacteria were suspended in tissue culture medium (TC) or mixtures of TC with BM or FF and applied to the E12 cultures. Bacterial numbers were assessed by fluorescence. DyLight 650-labelled neutravidin, which cannot cross cell membrane, evaluated extracellular bacteria. Fluorescence of basolateral medium was measured to quantify translocation. Bacterial concentrations were compared using the Mann Whitney U test. RESULTS: After 1h exposure, E12 cultures adhered or internalised more NEC-derived bacteria than standard strain E. coli and more suspended in FF than BM (P<0.001). A greater proportion of NEC-derived bacteria internalised when suspended in TC or BM. In FF, the NEC-derived strain internalised least. More translocation occurred in BM incubations compared to FF in the first 1-4h: NEC-E. coli less than the reference strain. After 24h translocated bacterial populations were equal. CONCLUSION: In this pilot study, breast milk was associated with relatively less adhesion and internalisation of NEC-associated E. coli to mucus covered E12s compared to formula milk.
Subject(s)
Bacterial Physiological Phenomena , Escherichia coli/physiology , Intestines/cytology , Milk, Human , Cells, Cultured , Enterocolitis, Necrotizing/microbiology , HT29 Cells , Humans , Pilot ProjectsABSTRACT
Turnover of mucins in supramucosal gels is essential for the removal of surface contaminants, and the maintenance of normal mucosal barrier function. In addition to the well-known processes promoting the physical turnover of mucus gels, extracellular mucin degradation also requires the coordinated action of a range of enzyme activities including glycosidases and proteases. These are collectively termed "mucinase". Derangements of mucinase activity lead to downstream barrier defects and mucosal disease. This chapter is focussed on methods that can be used to assess the degradation of whole mucins and isolated mucin glycans. A range of approaches is described using labelled or unlabelled substrates utilised in assays based on 96-well plates, size exclusion chromatography, and NP-HPLC. These are suitable for defining the extent and progress of mucin degradation in different mucosal systems, and identifying abnormalities and critical control points.
Subject(s)
Mucins/analysis , Mucins/metabolism , Animals , Humans , Polysaccharide-Lyases/metabolismABSTRACT
DMBT1 (deleted in malignant brain tumor 1), a human mucin-like glycoprotein, belonging to the scavenger receptor cysteine-rich (SRCR) superfamily, is mainly secreted from mucosal epithelia. It has been shown previously that interaction of hensin, the rabbit ortholog of DMBT1, with galectin 3, a ß-galactoside-binding lectin, induces a terminal differentiation of epithelial cells. In this paper, we have used surface plasmon resonance (SPR), to analyse the binding of galectin 3 to two purified samples of human DMBT1:recombinant DMBT1 produced in CHO cells and DMBT1 isolated from intestinal tissues. Characterization of their glycosylation profile by nano-ESI-Q-TOF tandem mass spectrometry showed significant differences in O-glycans between the two DMBT1 samples. Results obtained by SPR demonstrated that the oligosaccharide side chains of DMBT1 are recognized by the carbohydrate-recognition domain (CRD) of galectin 3 and modification in the pattern of oligosaccharides modulates the binding parameters of DMBT1 with galectin 3. Moreover, using immunohistochemistry on paraffin-embedded colonic tissue sections, we could show a co-localisation of DMBT1 and galectin 3 in human intestine, suggesting a potential physiological interaction.