Biochemical differences between trail mucus and adhesive mucus from marsh periwinkle snails

The composition of the adhesive form of marsh periwinkle mucus was compared to the trail mucus used during locomotion. The trail mucus consists primarily of large, carbohydrate-rich molecules with some relatively small proteins. In contrast, the adhesive mucus has 2.7 times as much protein with no significant difference in carbohydrate concentration. The resulting gel has roughly equal amounts of protein and carbohydrate. This substantial increase in protein content is due to the additional presence of two proteins with molecular weights of 41 and 36 kD. These two proteins are absent from the trail mucus. Both proteins are glycosylated, have similar amino acid compositions, and have isoelectric points of 4.75. This change in composition corresponds to an order of magnitude increase in tenacity with little clear change in overall concentration. The difference between adhesive and non-adhesive mucus suggests that relatively small proteins are important for controlling the mechanics of periwinkle mucus.     

Cyclic AMP in cervical mucus

Cyclic adenosine monophosphate normally stimulates motility of spermatozoa. Its concentration in cervical mucus was studied by an isotopic competitive method in 15 normal women aged between 20 and 50 years. Values were very high, particularly in the periovulatory period, with a mean (+/-SD) value of 167.90 +/- 154.96 nmol/l. These are very high when compared with values in other biological fluids (blood serum and urine).     

Microbial adhesins to gastrointestinal mucus

The gastrointestinal tract (GIT) is lined by a layer of mucus formed by mucin glycoproteins. This layer constitutes a physical and chemical barrier between the intestinal contents and the underlying epithelia. In addition to this protective role, mucins harbor glycan-rich domains that provide preferential binding sites for pathogens and commensal bacteria. Although mucus-microbial interactions in the GIT play a crucial role in determining the outcome of relationships of both commensal and pathogens with the host, the adhesins and ligands involved in the interaction are poorly delineated. This review focuses on the current knowledge of microbial adhesins to gastrointestinal mucus and mucus components.     

Enumerating viruses in coral mucus

The distribution of viruses inhabiting the coral mucus remains undetermined, as there is no suitable standardized procedure for their separation from this organic matrix, principally owing to its viscosity and autofluorescence. Seven protocols were tested, and the most efficient separations were obtained from a chemical treatment requiring potassium citrate.     

Selective permeability of mucus barriers

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Neuraminidase in pig gastric mucus

This paper demostrates the presence of neuraminidase (EC 3.2.1.18) in the isolated gastic mucoproteins, and indicates its probable origin from bacteria in the mucous secretion. The sialic acid content of gastric mucoproteins is discussed in terms of these findings.     

Biochemical and rheological analysis of human colonic culture mucus reveals similarity to gut mucus

The goal of this project was to validate the functional relevance and utility of mucus produced by an in vitro intestinal cell culture model. This is facilitated by the need to physiologically replicate both healthy and abnormal mucus conditions from native intestinal tissue, where mucus properties have been connected to intestinal disease models. Mucus harvested from colonic cell cultures derived from healthy donors was compared to mucus collected from surgically resected, noninflamed transverse colon tissue. The rheological and biochemical properties of these mucus samples were compared using oscillational rheometry, particle-tracking microrheology, multiangle laser light scattering, refractometry, and immunohistochemical imaging. An air-liquid interface culture of primary human colonic epithelial cells generated a continuous monolayer with an attached mucus layer that displayed increasing weight percent (wt%) of solids over 1 week (1.3 ± 0.5% at 2 days vs. 2.4 ± 0.3% at 7 days). The full range of mucus concentrations (0.9–3.3%) observed during culture was comparable to that displayed by ex vivo mucus (1.3–1.9%). Bulk rheological measurements displayed similar wt%-based complex viscosities between in vitro and ex vivo mucus, with the complex viscosity of both systems increasing with wt% of solids. Particle-tracking microrheology showed higher complex viscosities for ex vivo mucus samples than in vitro mucus which was explained by a greater fraction of water present in in vitro mucus than ex vivo, i.e., in vitro mucus is more heterogeneous than ex vivo. Refractometry, multiangle laser light scattering, and immunostaining showed increased mucus complex size in ex vivo mucus compared with in vitro mucus, which may have been due to the admixture of mucus and cellular debris during ex vivo mucus collection. The air-liquid interface culture system produced intestinal mucus with similar composition and rheology to native human gut mucus, providing a platform to analyze pathological differences in intestinal mucus.     

Anti-adhesive glycoproteins in echinoderm mucus secretions

Marine invertebrates produce a large variety of mucus secretions which are rich in glycoproteins. As part of our studies of natural antifouling mechanisms, mucus secretions from the starfish Marthasterias glacialis and Porania pulvillus and the brittlestar Ophiocomina nigra have been used to characterise the structure and function of some of the glycoproteins present in these secretions. Mucus was collected from all three species and fractionated by size exclusion chromatography. A high molecular weight glycoprotein fraction was collected from each species. Monosaccharide analysis and FTIR demonstrated a composition consistent with a mucin-type glycoprotein. The mucin from M. glacialis and O. nigra inhibited in vitro bacterial adhesion in a dose-dependent manner. In contrast, the mucin from P. pulvillus promoted bacterial adhesion in a dose-dependent manner. All of the mucins inhibited the adhesion of human neutrophils to cultured human vascular endothelial cells (HUVECs) and had no anticoagulant activity. The mucins described here have adhesion-regulating functions that may have a role in the antifouling or feeding mechanisms of the organisms that produce them. These mucins may also be of therapeutic value through their ability to regulate human neutrophil adhesion or bacterial adhesion.     

Biochemical examination ofRana pipiens epithelial mucus

Summary The epithelial mucus ofRana pipiens is shown to be highly negative by histochemical procedures, uptake of tagged extracellular markers, equilibrium dialysis, and QAE-Sephadex G-25 ion exchange chromatography. The mucus is found to contain 0.4% (dry weight) sulfate, 16% (dry weight) protein and 9% (dry weight) neutral sugars, whereas no sialic acid is detected. A mucin charge of –40 equivalents mole^–1 is calculated by equilibrium dialysis using a molecular mass of 100,000 Dalton. An independent determination of in situ charge density by radionuclide uptake onto the frog surface (1.55 meq l^–1) suggests that this estimate of mucin charge is correct. Based on analysis of size-exclusion chromatography fractions, it is suggested that mucus contains a sulfated glyco-protein (100,000 Dalton) that forms aggregates of about one million Dalton.     

A model for mucus glycoprotein structure

An approximately 500,000Da subunit characterizes the structural glycoprotein of mucus. This unit is composed of a rod-like very heavily glycosylated protein chain with an unglycosylated cysteine rich region at its end. It is proposed that the 'bare' peptide portion of the subunit forms itself into a lectin by undergoing a disulfide bond stabilised conformational fold. The lectin binding site, R, is assumed to be selective for a rare sugar sequence, which, when present, creates a binding site R. The site R has to be relatively rare. The average number less than s greater than of sidechains which contain R per subunit will only be of order 1. When less than s greater than less than or equal to 1 reasonably long, chain-like aggregates are formed which behave like coiling polymer chains with the subunit the Kuhn statistical element. When less than s greater than much greater than 1 the entire system forms one structure. Data obtained from the literature are analysed. They favor finite size, separate chains. The Kuhn statistical element length, derived from the data, is shown to agree well with the lectin model hypothesis.     

Coarse-grained modeling of mucus barrier properties

We designed a simple coarse-grained model of the glycocalyx layer, or adhesive mucus layer (AML), covered by mucus gel (luminal mucus layer) using a polymer lattice model and stochastic sampling (replica exchange Monte Carlo) for canonical ensemble simulations. We assumed that mucin MUC16 is responsible for the structural properties of the AML. Other mucins that are much smaller in size and less relevant for layer structure formation were not included. We further assumed that the system was in quasi-equilibrium. For systems with surface coverage and concentrations of model mucins mimicking physiological conditions, we determined the equilibrium distribution of inert nanoparticles within the mucus layers using an efficient replica exchange Monte Carlo sampling procedure. The results show that the two mucus layers penetrate each other only marginally, and the bilayer imposes a strong barrier for nanoparticles, with the AML layer playing a crucial role in the mucus barrier.