Interactions between Yersinia enterocolitica and purified intestinal mucins from rabbit and humans were investigated. Plasmid-bearing virulent organisms (but not plasmid-free nonvirulent bacteria) bound well to both mucins, suggesting that adherence was controlled by the virulence plasmid. Examination of binding to 14 different preparations of purified human intestinal mucin (8 preparations obtained from normal subjects and 6 samples from patients with cystic fibrosis) revealed no differences between normal and cystic fibrotic mucins in ability to serve as a binding substrate for virulent Y. enterocolitica. Analyses of binding curves suggested the presence of a single type of noninteracting receptor for Y. enterocolitica in both rabbit and human mucins with similar (but not necessarily identical) structures. Virulent bacteria bound to polystyrene through hydrophobic interactions that could be disrupted by treating the organisms with tetramethyl urea. In contrast, binding of plasmid-bearing Y. enterocolitica to intestinal mucin was not susceptible to tetramethyl urea and therefore does not appear to involve hydrophobic interactions. Prior incubation of organisms with mucin significantly inhibited binding to polystyrene, suggesting that mucin can mask hydrophobic adhesins on the bacterial surface. Hapten inhibition studies revealed that the monosaccharides galactose and N-acetylgalactosamine and the disaccharide lactose could markedly reduce (but not abolish) bacterial adherence to mucin but other monosaccharides and the RGD peptide had no effect on mucin binding. We conclude that virulent Y. enterocolitica is capable of interacting with the carbohydrate moiety of intestinal mucin. These interactions appear to be plasmid mediated and not hydrophobic.Yersinia enterocolitica is an enteroinvasive bacterium that causes gastroenteritis (4, 40). The mechanisms enabling the organism to colonize the gut and produce disease have not yet been clearly defined, but all pathogenic strains (in contrast to nonpathogenic environmental isolates) contain a 42to 50-MDa plasmid (6, 31), indicating that the plasmid is essential for virulence. A variety of proteins (16 to 20) are encoded by the plasmid, and their expression is regulated by both temperature and the availability of calcium (6,31,32,43). Some of these proteins are secreted, and a number then associate with the outer membrane of the organism and are referred to as Yops (for yersinia outer membrane proteins) (22,24,25,31,32). In addition, the plasmid encodes for YadA, a fibrillar adhesin that is a true outer membrane protein (12). When expressed, these plasmid-encoded proteins change the surface charge and hydrophobicity of the bacterium, promote autoagglutination and mannose-resistant hemagglutination, enhance adherence to cell lines, collagen, and fibronectin, and confer on the organism the ability to resist phagocytosis by polymorphonuclear leukocytes and the bacteriocidal effects of serum (1, 9, 12-15, 22, 23, 29, 41, 46). Thus, plasmid proteins may participate in attac...

Plasmid-bearing (but not plasmid-cured) Yersinia enterocolitica is known to bind to purified small intestinal mucins from rabbits and humans. This study examined which region(s) of the mucin molecule is important for bacterial adherence. Pronase digestion of mucin and removal of nonglycosylated or poorly glycosylated peptide regions had no effect on bacterial binding, suggesting that plasmid-bearing Y. enterocolitica interacts with mucin carbohydrate. Periodate oxidation also did not alter bacterial adherence, indicating that vicinal hydroxyl groups in the mucin sugars are not important for binding. Boiling of mucin, depolymerization by reduction of disulfide bonds, or removal of noncovalently associated lipid actually enhanced bacterial adherence, suggesting that plasmid-bearing Y. enterocolitica can interact with additional domains in the mucin molecule revealed by these treatments. These domains were destroyed by pronase digestion. In delipidated mucin (but not in reduced or boiled mucin), binding to these domains appeared to be hydrophobic since it could be prevented by treatment of bacteria with tetramethyl urea. Oligosaccharides obtained from both human and rabbit small intestinal mucins were capable of inhibiting attachment of plasmid-bearing (but not plasmid-cured) Y. enterocolitica to mucin. After removal of terminal and backbone sugar residues by treatment of mucin with trifluoromethanesulfonic acid, binding of plasmid-bearing bacteria increased significantly when N-acetylgalactosamine, either alone or with galactose attached, was revealed, indicating that core regions of the sugar side chains are involved in bacterial binding. Adherence of plasmid-cured organisms was unaffected by trifluoromethanesulfonic acid treatment of mucin. We concluded that virulent Y. enterocolitica interacts with the carbohydrate moiety of native small intestinal mucin through a plasmid-mediated process. When mucin becomes denatured, binding of the organism can increase through hydrophobic and nonhydrophobic interactions with (most likely) the mucin protein.

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