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Moist mucosal epithelial interfaces that are exposed to external environments are dominated by sugar epitopes, some of which (e.g., sialic acids) are involved in host defense. In this study, we determined the abundance and distribution of two sialic acids to assess differences in their availability to an exogenous probe in isolated mucins and mucous gels. We used atomic force microscopy to obtain force maps of human preocular mucous and purified ocular mucins by probing and locating the interactions between tip-tethered lectins Maackia amurensis and Sambucus nigra and their respective receptors, α-2,3 and α-2,6 N-acetylneuraminic (sialic) acids. The rupture force distributions were not affected by neighboring sugar-bearing molecules. Energy contours for both lectin-sugar bonds were fitted to a two-barrier model, suggesting a conformational change before dissociation. In contrast to data from purified mucin molecules, the preocular gels presented numerous large clusters (19,000 ± 4000 nm(2)) of α-2,6 sialic acids, but very few small clusters (2000 ± 500 nm(2)) of α-2,3 epitopes. This indicates that mucins, which are rich in α-2,3 sialic acids, are only partially exposed at the surface of the mucous gel. Microorganisms that recognize α-2,3 sialic acids will encounter only isolated ligands, and the adhesion of other microorganisms will be enhanced by large islands of neighboring α-2,6 sialic acids. We have unveiled an additional level of mucosal surface heterogeneity, specifically in the distribution of pro- and antiadhesive sialic acids that protect underlying epithelia from viruses and bacteria., (Copyright © 2012 Biophysical Society. Published by Elsevier Inc. All rights reserved.)

High-resolution atomic force microscopy has been applied to the imaging of intact human ocular mucins in a near-physiological buffer. The mucins displayed a range of lengths from several hundred nanometers to several microns. By varying the ionic composition of the imaging environment, it was possible to image molecules rigidly fixed to the substrate and the motion of single molecules across the substrate. From static molecular images, high-resolution line profiles show a variation of up to +/-0.75 nm in thickness along the molecule. This variation is localized in regions of several tens of nanometers. It is interpreted in terms of the varying glycosylation along the mucin and is consistent with the known size of oligosaccharides in ocular mucins. The dynamic images indicate the possibility of following mucin interactions in situ.